HK40005642A - Novel cyclosporin derivatives and uses thereof - Google Patents

Description

Novel cyclosporin derivatives and use thereof

RELATED APPLICATIONS

This application is related to U.S. application serial No. 13/840,088 filed on 3, 15, 2013. This application claims priority to U.S. provisional application No. 62/337,377 filed on day 17/5/2016, U.S. provisional application No. 62/339,464 filed on day 20/5/2016, and U.S. provisional application No. 62/384,822 filed on day 8/9/2016. The entire contents of these applications are expressly incorporated herein in their entirety by reference.

Technical Field

The present invention relates to novel cyclosporin derivatives, pharmaceutical compositions containing them, and methods for treating or preventing viral infections, inflammation, dry eye, central nervous disorders, liver, lung and kidney diseases, cardiovascular diseases, cancer, obesity, diabetes, muscular dystrophy, alopecia, and the like.

Background

Cyclosporin is essentially a poly-N-methyl cyclic undecenoic acid polypeptide isolated from fungi. Cyclosporin a has immunosuppressive activity and has been used for nearly 30 years to prevent rejection in kidney, heart and liver transplant recipients. It has anti-inflammatory properties and has been used to treat severe rheumatoid arthritis, severe psoriasis, Behget uveitis and dry eye. In addition, it can be used for the treatment of severe ulcerative colitis, Crohn's disease, alopecia areata, aplastic anemia, HSV-1 stromal keratitis, systemic lupus erythematosus and severe lupus nephritis. However, its strong immunosuppressive activity limits its use in many diseases.

The anti-HIV activity of cyclosporin A was first discovered in 1986 and has been studied since then (Klatzmann, D., et al, 1986, C R Acad. Sci.III,303(9): 343-8; Wainberg, M.A., et al, 1988, Blood,72,1904-10; Luban, J., et al, 1993, Cell,73, 1067-one 1078, each of which is incorporated herein by reference). Its non-immunosuppressive derivative, NIM-811, is reported to have potent anti-HIV activity because it is capable of inhibiting cyclophilin A (Franke, E.K., et al, 1994, Nature,372, 359-containing 362; Thali, M., et al, 1994, Nature,372, 363-365; Gamble, T.R., et al, 1996, Cell,87, 1157-containing 1159; Rosenwirth B., et al, 1994, Antiicrob. Agents Chemother, 38, 1763-containing 1772; each of which is incorporated herein by reference).

Cyclosporin A and its non-immunosuppressive derivatives, such as NIM-811 (N-MeIle-4-cyclosporin), Debio-025, SCY-635, EDP-494, DEP-546, NIM-258, CPI-431-32(CRV431), and STG-175, bind to and inhibit cyclophilins, subsequently preventing HCV RNA replication and protein synthesis. As a result, these compounds have potent anti-HCV activity (Watashi, K., et al, 2007, Rev. Med. Virol.,17: 245-252.37; Inoue, K., et al, 2001, Nippon Rinsho.,59,1326-30; Inoue, K., et al, 2003, J.Gastroenterol.,38,567-72; Watashi, K., et al, 2003, Hepatology,38,1282-8; Gaither, L.A., et al, 2010, Virology,397, 43-55; Rhodin, M.H.J., et al, 2016, EASL, Poster 250, and the news of the 34 (see)^thAnnual l.p. morgan Healthcare, Conference Presentation on jan.13,2016at 11:00AM byEnanta pharmaceutical.inc. (www.enanta.com) (EDP-494); baugh, J.M., et al, 2013, Antiviral Res.100(2): 555-; fu, j., et al, 2014, j.med.chem.,57, 8503-; gallay, P.A., et al, 2015, PLoS One 10(8), e0134707 doi 10.1371/journal. po. 0134707(CPI-431-32(CRV 431)); gallay, P.A., et al, 2016, PLoS One11(4): e0152036.d10.1371/journal. bone.0152036 (STG-175); each of which is incorporated herein by reference). NIM-811, Debio-025 and SCY-635 have been evaluated in phase II and III clinical trials against HCV, and the phase I EDP-494 study is beginning in 2016 for resistance-related variants of HCV in the first quarter.

NIM-811 and Debio-025 have chemical structures similar to cyclosporin a and have poor pharmacokinetic properties. In addition, they are metabolized by P450, which induces drug interactions (Lill, J., et al, 2000, Am J Health-SystPharm 57,1579; incorporated herein by reference).

SCY-635 has an improved pharmacokinetic profile and low serum binding. In addition, it has low potential for drug-drug interactions. The in vitro anti-HCV activity (EC50) of SCY-635 was reported to be 0.10. mu.M (Hopkins, S. et al, 2010, Antimicrob. Agents Chemother.,54,660-672, incorporated herein by reference). However, SCY-635 is not chemically stable, since it is readily converted to its diastereomer by epimerization. Its diastereomers are expected to have poor binding activity to cyclophilins, and as a result, its antiviral activity in vivo may be affected (see, e.g., WO2012/009715, WO2012/021796, and WO2012/075494, each of which is incorporated herein by reference in its entirety).

Cyclosporin a and its non-immunosuppressive derivatives were also found to have anti-HBV activity through inhibition by cyclophilins (Chokshi, s., et al, 2012, but 61: a 11; Chokshi, s., et al, 2012, post Presentations,47th Annual Meeting of the European Association for the Study of the lever (EASL 2012), Barcelona, Spain; Chokshi, s., et al, 2011, Abstract 190 (postpresentations), 46th Annual Meeting of the European Association for the Study of the light over (EASL 2011), Berlin, ch 30-April 3; tiana, x.c., et al, 2010, j.virol, 84,3373, xl 3381, xhol, w.771, 2003, g., 9, p.18, p.7, p., p.3; each incorporated herein by reference, p.3, Michael, p.3, h.

In addition, cyclophilins are reported to regulate the life cycle and pathogenesis of several viruses, including severe acute respiratory syndrome coronavirus, vaccinia virus and herpes simplex virus (Castro, A.P., et al, 2003, J.Virol.,77, 9052-. Cyclosporin a and its non-immunosuppressive derivatives also have such antiviral activity.

N-MeVal-4-cyclosporin (SDZ 220-384), another non-immunosuppressive cyclosporin derivative, has been reported to have biological activity similar to that of NIM-811 (Fliri, H., et al, 1993, Ann. N Y Acad Sci.696, 47-53; Zenke, G., et al, 1993, Ann N Y Acad Sci.23; 685: 330-5).

Hepatitis C Virus (HCV) is a small (55-65 nm in size) enveloped positive-sense single-stranded RNA virus in the Flaviviridae family. HCV has a high replication rate and a particularly high mutation rate. Approximately 80% of people infected with HCV develop chronic persistent infections. More than 400 million americans have been infected with HCV and it is estimated that more than 200 million people are chronically infected worldwide. It is estimated that about 35,000 new cases of hepatitis c occur in the united states each year. In the united states, HCV infection causes approximately 50% of the chronic liver disease, 30% of all liver transplants, and 30% of all liver cirrhosis, end stage liver disease, and liver cancer. peg-interferon and ribavirin combinations are the standard therapy for chronic hepatitis c, but have low efficacy against HCV infection. Recently, the FDA has approved Vertex's inclik (telaprevin) and Merck's Victrelis (boceprevin) as adjuncts to current interferon/ribavirin therapies for the treatment of HCV. Both of these drugs are HCV protease inhibitors that target the virus to prevent its replication. However, due to the rapid mutation rate of HCV, resistance can develop in a short period of time. Thus, there is a need for an effective therapeutic agent for HCV treatment.

Hepatitis B Virus (HBV) is a 42nm partially double-stranded DNA virus consisting of a 27nm nucleocapsid core (HBcAg) surrounded by an outer lipoprotein envelope containing surface antigen (HBsAg). more than 2 hundred million people are infected, and there are 3.5 million chronic virus carriers which have caused epidemic conditions in parts of Asia and Africa. chronic hepatitis B causes cirrhosis and liver cancer, a fatal disease that is not very responsive to current chemotherapy.the infection can be prevented by vaccination and can be reduced by current antiviral drugs such as lamivudine (Epivor), adefovir (Hepsera), tenofovir (Viread), tebufenvudine (Tyzeka), entecavir (Baraclude), and two immune system modulators interferon α -2a and pegylated interferon α -2a (Pegasys) (HBV 14, 31. A, 31. J.8, 14. A, 14. 10, 14. J.8. A, 14. A, 14, 5, g, 5. A, J. 14, 5, J. A, 5.

Non-immunosuppressive cyclosporin derivatives bind to cyclophilins, a family of peptidyl-prolyl cis-trans isomeric host proteins that catalyze protein folding and regulation, critical for the processing and maturation of viral proteins for viral replication. HIV and HCV are high mutation rate viruses. All current antiviral drugs target the virus itself; when the virus becomes virulent, it leads to the development of resistance. Targeting host cofactors (cyclophilins) will slow the development of Resistance due to a higher genetic barrier, not directly to the virus (Rosenwirth, B., et al, 1994, anti. organic chemistry, 38, 1763-1772; Tang, H.L. et al, 2010, Virus, 2, 1621-1634; Hopkins, S. et al, 2010, Oral Presentation, Scanexis's SCY-635 expression barrierresistance in HCV Treatment, the 45th annular Meet of the European Association for the Study of the Liver (EASL 2010), Vienna, stria, April 14-18; each of which is incorporated herein by reference). Cyclosporin derivatives affect a new target, the cyclophilins, and thus represent a new mechanism of action against viruses.

There are 19 cyclophilins in the human genome (Thapar, r.,2015, biomodules 5(2):974-99.), but the function of these cyclophilin isoforms remains unclear (Davis, t.l., et al, 2010, PLoS biol.8(7): e 1000439; incorporated herein by reference). Cyclophilins A, B, C, D and other such isoforms play an important role in the pathophysiology of many serious diseases, such as Cancer (Campa, MJ., et al, 2003, Cancer Res.,63(7), 1652-6; Li, M., et al, 2006, Cancer,106: 2284-94; Yang, H., et al, 2007, Biochem Biophys Rescomm., 361(3): 763-7; Obchoei, S., et al, 2009, Med Sci.,15 (11), RA 221-32; Andersson, Y., et al, 2009, Br J Cancer,101,1307-, 12: 442; lee, j.,2010, arch, pharm. res.,33(9) 1401-9; hamilton, G.,2014, curr. cancer Drug Target,14(1): 46-58; zhu, d., et al, 2015, nat. med.,21(6) 572-80; lavin, p.t., et al 2015, curr.mol.pharmacol, 9(2) 148-64; seleh, t, et al, 2016, nat. chem.biol.,12(2): 117-23; each of which is incorporated herein by reference), inflammation (the result of interactions between secreted extracellular cyclophilins and CD-147 (surface protein); yurchenko v.,2005, Immunology,117(3): 301-9; yurchenko, v.,2010, Clin Exp immunol.,160(3): 305-17;m. 2010, Angew Chem Int Ed Engl, 49(1): 213-5; each of which is incorporated herein by reference), cardiovascular disease (including blood vessels)Stenosis, atherosclerosis, abdominal aortic aneurysm, aortic rupture, cardiac hypertrophy, pulmonary hypertension, myocarditis and myocardial fibrosis and ischemic heart disease, liver, kidney and pulmonary fibrosis, protection and regeneration; jin, z.g., et al, 2000, Circ res, 87(9): 789-96; yurchenko, v., et al, 2005, Immunology,117, 301-; suzuki, J., et al, 2006, Circ Res, 98(6): 811-7; satoh, k., et al, 2008, circulation,117 (24): 3088-98; nishihara, M., et al, 2008, J Mol Cell Cardiol, 44(2) 441-; satoh, k, et al, 2010, Circ j, 74(11) 2249-56; satoh, k., et al, 2010, artificial Redox signal, 12(5) 675-82; hausenloy, D.J., et al, 2012, Br JPharmacol.165(5): 1235-45; coppinger, J.A., et al, 2004, Blood,103(6): 2096-; satoh, K., et al, 2010, Artificial Redox Signal, 1:12(5), 675-682; nigro, p., et al, 2010, J Exp med.,208(1): 53-66; wang, W.L., et al, 2011, Med Hypotheses,77(5): 734-8; hattori, f.,2012, J Mol Cell cardio, 53(1): 1-2; seizer P.,2012, J Mol CellCardiol, 53(1): 6-14; naoumov, n.v.,2014, l.hepatol.,61(5) 1166-74; each of which is incorporated herein by reference), rheumatoid arthritis (Wells, g., et al, 2000, Cochrane Database systrev., (2): CD 001083; kim, h, et al, 2005, Clin immunol, 116(3): 217-24; yang, Y., Rheumatology (Oxford),47(9), 1299-; yurchenko, v., et al, 2006, Immunology,117(3): 301-9; damsker, j.m.,2009, Immunology,126(1): 55-62; wang, L., et al, 2010, J Clin Immunol.,30(1): 24-33; billch A., et al, 1997, J Exp Med.,185: 975-80; de CeuninckF, et al, 2003, Arthritis Rheum, 48: 2197-; each of which is incorporated herein by reference), respiratory inflammation (Foda, h.d., et al, 2001, Am J Respir Cell Mol biol.,25: 717-24; hasaneen, N.A., et al, FASEB J.,19:1507-9.Yurchenko, V.et al, 2006, Immunology,117(3): 301-9; gwinn, w.m.,2006, J immunol, 177(7): 4870-9; onoue, S.,2009, J Control Release, 138(1): 16-23; balsley, m.a., et al, 2010, J immunol.,185(12) 7663-70; balsley, m., et al, 2010, am.j.respir.crit.care med.,181(1): a 6821; stemmy, e.j., et al, 2011, j.asthma,48(10): 986-; stemmy, e.j., et al, 2011, Am J Respir Cell Mol biol.,45(5): 991-8; amin, k.,2012, Respir med.,106(1): 9-14; one oue, S.,2012, Eur J Pharm Biopharm.,80(1) 54-60; each of which is incorporated herein by reference), lupus (cccavo, d., et al, 1997, Arthritis)&Rheumatosis, 40(1) 27-35; dost l, C, et al, 1998, Lupus,7(1): 129-36; tam, LS., et al, 1998, Q J Med.,91(8): 573-580; fu, LW., et al, 1998, Rheumatology 37(2): 217-; hallegua, D., et al, 2009, Lupus,9: 241-; each of which is incorporated herein by reference), psoriasis (Ellis, c.n.,1991, N Engl J med, 324, 277-; lebwohl, M., et al, 1998, J Am AcadDermatol.,39(3): 464-75; roscarin, DM., et al, 2010, J Am Acad German, 62(5): 838-53; each of which is incorporated herein by reference), atopic dermatitis (Naeyaert, j.m., et al, 1999, Dermatology,198: 145-; pacor, ML., et al, 2001, Recenti Prog med.,92(6): 390-1; ricci, G., et al, 2009, Drugs,69(3):297 oz 306; simon, D.,2011, Curr Probl German, 41: 156-64; each of which is incorporated herein by reference), dry eye disease (Pflugfelder, s.c.,2004, Am jophthalmol, 137(2), 337-42; kymionis, g.d., et al 2008, Clin ophthalmol, 2, 829-836; kunert, k.s., et al, 2002, Arch ophthalmol, 120,330-7; yavuz, B., et al, 2012, Scientific World journal.2012: 194848; each of which is incorporated herein by reference), severe Graves eye disease (Prummel, m.f.,1989, N Engl J med.,321(20), 1353-9; incorporated herein by reference), endogenous uveitis (Nussenblatt, r.b., et al, 1991, Am J ophthalmol, 112(2), 138-46; incorporated herein by reference), wegener's granulomatosis (georgana, c., et al, 1996, Clin rheumato, 15(2), 189-92; incorporated herein by reference), vernal keratoconjunctivitis (Pucci, n., et al, 2002, Ann Allergy asthma immunol.,89, 298-; incorporated herein by reference), atopic keratoconjunctivitis (Akpek, e.k., et al, 2004, Ophthalmology,111,476-82; incorporated herein by reference), wood-like conjunctivitis (Rubin, b.i., et al, 1991, AmJ ophthalmol.,112, 95-96; incorporated herein by reference), conjunctival lichen planus (Levell, n.j., et al, 1992, Br J dermatol.,127, 66-7; incorporated herein by reference), superior limbal keratoconjunctivitis (Perry, h.d., et al, 2003, Ophthalmology,110,1578-81; incorporated herein by reference), inflammatory Bowel disease-crohn's disease and ulcerative colitis (Sandborn, w.j.,1995, inflam Bowel dis.1: 48-63;shibolet, O., et al, 2005, Cochrane Database Syst Rev., (1): CD 004277; rufo, p.a., et al, 2006, Paediatr Drugs,8(5): 279-302; reindl, w., et al, 2007, gut, 56(7): 1019; hart, a.l., et al, 2010, AlimentPharmacol ther, 32(5) 615-27; cheifetz, a.s., et al, 2011, J Clin gastroenterol.,45(2) 107-12; sharkey, L.,2011, J Crohns Colitis, 5(2): 91-4; fabro, M., et al, 2011, Curr Drug targets, 12(10): 1448-53; van Assche, g., et al, 2011, gut, 60(1): 130-3; each of which is incorporated herein by reference), NSAID-induced bowel disease (louguidic, a., at al.,2010, toxicol. sci.,118, 276-; which is incorporated herein by reference) and ischemic brain diseases (Boulos, s., et al, 2007, Neurobiol dis.,25: 54-64; incorporated herein by reference).

Due to the cyclophilin inhibition, cyclosporin derivatives also possess the following biological activities: antifungal (Kirkland, T.N., et al, 1983, Antirhizob Agents Chemother.,24(6): 921-924; Mody, C.H., et al, 1988, infection Immun.,56(1): 7-12; Roilides, E.et al, 1994, Antirhizob Agents Chemother.,38(12): 2883-2888;m. et al 1997, Appl Environ Microbiol.,63(5): 1739-43; cruz, M.C., et al, 2000, Antimicrob Agents Chemother, 44(1) 143-9; each of which is incorporated herein by reference), antimalarial (nichell, s.p., et al, 1982, infection immun, 37(3): 1093-; murphy, J.R., et al, 1988, Antimicrob Agents Chemother, 32(4) 462-6; mari i n-Men endez, A., et al, 2012, Mol Biochem Parasitol, 184(1): 44-7; each of which is incorporated herein by reference) and antiparasitic agents (including Leishmania donovani, Cryptosporidium, Microthecium taenicum, Toxoplasma gondii, Trypanosoma cruzi and Schistosoma japonicum; chappell, L.H., et al, 1992, Parasitology,105Suppl: S25-40; bell, A., et al, 1996, Gen Pharmacol, 27(6): 963-71; yau, W.L., et al, 2010, PLoS Negl Trop Dis, 4(6) e 729; yurchenko, v., et al, 2008, Int J parasitol, 38(6) 633-9; perkins, M.E., et al, 1998, Antichronob Agents Chemother, 42(4): 843-8; matsuzawa, K., et al, 1998, Int JParasitol, 28(4): 579-88; silverman, J.A., et al, 1997, Antimicrob AgentsChemother, 41(9) 1859-66; b-a, J., et al, 2008, Parasitology,135(2): 217-28; b-a, J., et al, 2004, Bioorg Med Chem Lett.,14(18): 4633-7; bout, D.T, et al, 1984, Am J tropimed hyg, 33(1): 185-6; bout, d, et al, 1986, infection immun, 52(3): 823-7; munro, G.H., et al, 1991, Parasitology,102Pt 1: 57-63; each of which is incorporated herein by reference), and further, cyclosporin derivatives can promote hair growth (Watanabe, s., et al, 1991, J dermaltol, (12): 714-9; PausR, et al, 1994, J Invest Dermatol, 103:2,143-7; hozumi, Y., et al, 1994, J DermatolSci, 7Suppl:, S33-8; takahashi, T., et al, 2001, J Invest Dermatol.,117(3): 605-11; taylor M., et al, 1993, J Invest Dermatol.,100:3,237-9; the amount of the gas-Gvili,

a., et al, 2004, Arch Dermatol Res.,296(6): 265-9; each of which is incorporated herein by reference).

Studies on alzheimer's disease indicate that cyclophilin a is a key target for the treatment of APOE 4-mediated neurovascular injury and the resulting neuronal dysfunction and degeneration (Bell, r.d., et al, 2012, Nature,485(7399): 512-6; Bell, r.d., et al, 2009, Acta neuropathohol, 118(1): 103-13; each of which is incorporated herein by reference).

Due to the function of extracellular cyclophilins, it is important to emphasize that the use of specific targets of secreted extracellular cyclophilins of cell-impermeable derivatives of cyclosporine will be effective in reducing inflammation of diseases such as respiratory tract inflammation and cardiovascular disease (Yurchenko v.,2005, Immunology,117(3): 301-9; Yurchenko, v.,2010, Clin expemmunol, 160(3): 305-17;m. 2010, Angew Chem Int Ed Engl, 49(1): 213-5; balsley, m.a., et al, 2010, J immunol.,185(12) 7663-70; balsley, m., et al, 2010, am.j.respir.crit.care med.,181(1): a 6821; satoh, k, et al, 2010, Circ j, 74(11) 2249-56; bukrinsky, M.,2015, Biochim Biophys Acta,1850(10) 2087-95; each of which is incorporated herein by reference). To target extracellular parentsCyclins, MM284 has been found and tested for its anti-inflammatory properties (Malesevic, m., et al, 2013, j.med. chem.,56, 7302-7311.). further studies have been reported on Biliary Atresia (BA) and other chronic conditions in the liver (Iordanskaia, t., et al, 2015, mol.med.,21(1):657-664.), on reduction of myocardial inflammation and cardiac fibrosis (Heinzmann, d., et al, 2015, PLoS One 10(8): ee0124606.doi: 10.1371/journal. point. 4606), on reduction of TNF- α (diatkovski, m., et al, 2015, j.rmacol. exp.r. (353-5.).

Cyclophilin d (cypd) is important for mitochondria-related neurological and cardiovascular functions because it is an integral part of the mitochondrial permeability transition pore (mPTP). Unregulated opening of mPTP leads to mitochondrial swelling and cell death. Thus, the CypD-mediated mPTP is directly linked to a new drug treatment strategy for many neurological and cardiovascular diseases such as alzheimer's disease, parkinson's disease, huntington's disease, ALS, aging, heart failure, brain trauma, spinal cord injury, epilepsy, stroke, ischemia reperfusion injury in the brain, heart, liver, lung, kidney and especially in myocardial infarction. The CypD-mediated mPTP is also associated with a new therapeutic strategy for cancer, obesity, diabetes and muscular dystrophy, liver fibrosis, liver protection and regeneration (Henry-Mowatt, J.,2004, Oncogene,23,2850-60; Galluzzi, L.,2006, Oncogene,25, 4812-4830; Hirai, K., et al, 2001, J Neurosci, 21, 3017-containing 3023; Friberg, H., et al, 2002, Biochimie,84, 241-250; Waldmeier, P.C., 1712003, Curr Mem., 10, 1485-containing 506; Hansson, M.J., 2006, 2004, J Bioeneger, 36,407-13; Manllin, P.G., et al, Resosj, NeosJ., 85, Busy, H., 120, Brix H., 10, Brucer, H., 120, Brucer, J., 190, J., G., J., USA, No. 1713, No. 35, No. 11, No. C., Brucer, No. C., Brusson, No. C., Brussn, No. C., No. 10, No. C., 2006, No. C., No. 10, No. C., 2006, No. C, g., et al, 2007, Physiol rev.,87, 99-163; ibarra, a., et al, 2007, Brain res, 1149,200- "209; michelakis, e.d., et al, 2008, Circulation,117, 2431-; du, h., et al, 2008, Nature Medicine,14, 1097-; piot c, et al, 2008, NEngl J med, 359,473-81; hatton, J., et al, 2008, J neurosurg.,109, 699-707; tatsuta, t., et al, 2008, EMBO J,27, 306-; reutenauer, J., et al, 2008, Br J pharmacol, 155,574-84; mazzeo, a.t., et al, 2009, Exp neurol, 218, 363-; galluzzi, L., et al, 2009, Nature Rev Neurosci.,10, 481-494; halestrap, a.p., et al, 2009, Biochim biophysicaca, 1787,1402-15; arnett, a.l.h., et al, 2009, curr.opin.genet.dev.,19, 290-; tiecolo, t., et al, 2009, Br J pharmacol.,157, 1045-; lessing, e.r., et al, 2010, neurouscu disc, 20,753-60; halestrap, A.P., et al, 2010, Biochem Soc trains, 38, 841-860; cernak, i., et al, 2010, J Cereb Blood Flow metab, 30,255-66; elrod, J.W., et al, 2010, J Clin invest, 120, 3680-3687; duchen, m.r., et al, 2010, Essays biochem.,47,115-37; schapira, a.h.v., et al, 2011, Parkinson's Disease, Volume 2011,1-7Article ID 159160; osman, M.M., et al, 2011, neuropeps, 45, 359-; devalaja-narashima k, et al, 2011, FEBS lett, 585,677-82; fujimoto, K., et al, 2010, Proc NatlAcad Sci U S A.107, 10214-9; irwin, W.A., et al, 2003, Nat Genet, 35, 267-271; angelin, A., et al, 2007, Proc Natl Acad Sci U S A,104,991-6; merlini, l., et al, 2008, Proc Natl Acad Sci U S a,105,5225-9; millay, d.p.,2008, Nat med.,14,442-7; malonuitre, s., et al, 2009biochem.j.,425(1): 137-48; dear, j.w., et al, j.immunol.,187(6): 3347-52; naoumov, n.v.,2014, l.hepatol.,61(5) 1166-74; each of which is incorporated herein by reference). Cyclosporin a and its derivatives can block CypD to prevent mitochondrial swelling and cell death and therefore can be used in the treatment of the above diseases, for example, as neuroprotective and cardiovascular and hepatoprotective agents or as novel mitochondrial agents.

Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs. Although methods and materials similar or equivalent to those described herein can be used in the practice or testing of the present invention, suitable methods and materials are described below. All publications, patent applications, patents, and other references mentioned herein are incorporated by reference in their entirety. In addition, the materials, methods, and examples are illustrative only and not intended to be limiting.

Disclosure of Invention

In one aspect, the invention provides compounds of formula (I):

or a pharmaceutically acceptable salt thereof, wherein:

x is 0 or 1;

R₈is alkyl, alkenyl, alkynyl, cycloalkyl, cycloalkenyl, aryl or heteroaryl; wherein R is₈Is covered with

Or a plurality of R₁Substitution; provided that R is₈-R₁Is not n-butyl or (E) -but-2-enyl;

R₂is ethyl, 1-hydroxyethyl, isopropyl or n-propyl;

w is NR₁O, S or CH₂；

R₃Is H, alkyl or substituted alkyl, alkenyl or substituted alkenyl, alkynyl or substituted alkynyl, cycloalkyl or substituted cycloalkyl, cycloalkenyl or substituted cycloalkenyl, aryl or substituted aryl or heteroaryl or substituted heteroaryl; wherein R is₃Optionally substituted by one or more R₁Substitution;

R₁each occurrence is independently H, halogen, aryl OR substituted aryl, heteroaryl OR substituted heteroaryl, OR_A、SR_A、NR_AR_B、-NR_A(CH₂)_OOR_B、-N((CH₂)_OOR_A)((CH₂)_OOR_B)-C(＝O)R_A、-C(＝O)OR_A、-OC(＝O)R_A、-OC(＝O)(C₁-C₆alkyl-R_H)、-OC(＝O)(CH₂)_oOR_A、-C(＝O)NR_AR_B、-NR_AC(＝O)R_B、-N(C(＝O)R_A)(C(＝O)R_B)、-N(C(＝O)(C₁-C₆alkyl-R_H))(C(＝O)(C₁-C₆alkyl-R_H))、-N(C(＝O)(C₁-C₆alkyl-R_H))₂、-NR_AC(＝O)(CH₂)_OOR_B、-NR_AC(＝O)(CH₂)_OOR_B、-N(C(＝O)(CH₂)_OOR_B)₂、-NR_AC(＝O)(CH₂)_ONR_AR_B、-NR_A(CH₂)_OC(＝O)OR_B、-N((CH₂)_OC(＝O)OR_A)((CH₂)_OC(＝O)OR_B)、-NR_A(CH₂)_OC(＝O)NR_AR_B、-N((CH₂)_OC(＝O)NR_AR_B)((CH₂)_OC(＝O)NR_AR_B)、-NR_A(CH₂)_OC(＝O)NR_A(CH₂)_OOR_B、-C(＝O)N((CH₂)_OOR_B)₂、-N((CH₂)_OC(＝O)NR_A(CH₂)_OOR_B)((CH₂)_OC(＝O)NR_A(CH₂)_OOR_B)、-C(＝O)NR_A(CH₂)_OOR_B、-C(＝O)NR_A(CH₂)_OOR_B、-C(＝O)NR_A(CH₂)_ONR_AR_B、-N＝CR_A-NR_AR_B、-NR_B-C(＝NH)-NR_AR_B、O(CH₂)_mOR_A、O(CH₂)_mCOOR_A、O(CH₂)_mCONR_AR_B、O(CH₂)_mCONR_A(CH₂)_mOR_A、O(CH₂)_mO(CH₂)_mOR_A、O(CH₂)_mNR_AR_B、O(CH₂)_mO(CH₂)_mNR_AR_B、NR_C(CH₂)_mNR_AR_B、NR_C(CH₂)_mNR_C(CH₂)_mNR_AR_B、Wherein said aryl or heteroaryl is optionally substituted by a substituent selected from the group consisting of halogen, hydroxy, (C)₁-C₆) Alkyl group, (CH)₂)_pOR_A、(CH₂)_pNR_AR_B、(CH₂)_pC(＝O)NR_AR_BAnd (CH)₂)_pC(＝O)OR_AAnd may be the same or different and is substituted by one or more groups;

R₇is composed of

Each R₅Independently is H, alkyl or substituted alkyl, alkenyl or substituted alkenyl, alkynyl or substituted alkynyl, cycloalkyl or substituted cycloalkyl, cycloalkenyl or substituted cycloalkenyl or aryl or substituted aryl;

R_Aand R_BIndependently for each occurrence:

hydrogen;

(C₁-C₆) Alkyl, optionally substituted by one or more radicals R which may be the same or different_DSubstitution;

(C₂-C₆) Alkenyl or (C)₂-C₆) An alkynyl group;

optionally is (C)₁-C₆) Alkyl substituted (C)₃-C₇) A cycloalkyl group;

optionally substituted by a group selected from halogen, -O (C)₁-C₆) Alkyl, -C (═ O) O (C)₁-C₆) Phenyl or benzyl of alkyl, amino, alkylamino and dialkylamino, which may be substituted identically or differently by 1 to 5 radicals;

or may be a saturated or unsaturated heterocyclic ring containing 5 or 6 ring atoms and 1 to 3 heteroatoms, which may be the same or different, selected from nitrogen, sulphur and oxygen;

or R_AAnd R_BTogether with the nitrogen atom to which they are attached form a saturated or unsaturated heterocyclic ring containing from 3 to 7 ring atoms, which may optionally contain another heteroatom selected from nitrogen, oxygen and sulfur and may optionally be substituted by 1 to 4 groups, which may be the same or different, selected from alkyl, phenyl and benzyl;

R_Ceach occurrence is independently hydrogen or (C)₁-C₆) An alkyl group;

R_Dindependently at each occurrence is halogen, hydroxy, O (C)₁-C₄) Alkyl, C (═ O) (C)₁-C₄) Alkyl, C (═ O) O (C)₁-C₄) Alkyl orWherein Z is CH₂、O、S、NH、NCH₃NEt, N-isopropyl, N-neopentyl, N-CH₂CH₂OH or N-CH₂CH₂OMe；

R_GEach occurrence is independently R_A、OR_A、SR_A、NR_AR_B、–(CH₂)_oR_A、–(CH₂)_oC(＝O)OR_A、–(CH₂)_oC(＝O)NR_AR_B、C(＝O)OR_A、OC(＝O)R_A、NR_AC(＝O)R_B、NR_AC(＝O)(CH₂)_OOR_A、C(＝O)O(CH₂)_OOR_A、C(＝O)OR_B、C(＝O)NR_AR_B、C(＝O)NR_A(CH₂)_OOR_B、C(＝O)N((CH₂)_OOR_A)((CH₂)_OOR_B)、C(＝O)N((CH₂)_OC(＝O)OR_A)((CH₂)_OC(＝O)OR_B)、C(＝O)N((CH₂)_ONR_AR_B)((CH₂)_ONR_AR_B)、C(＝O)N((CH₂)_OOC(＝O)(CH₂)_OOR_A)((CH₂)_OOC(＝O)(CH₂)_OOR_B)、C(＝O)N((CH₂)_ONR_AC(＝O)(CH₂)_OOR_B)((CH₂)_ONR_AC(＝O)(CH₂)_OOR_B)、C(＝O)NR_A(CH₂)_ONR_AR_B、、C(＝O)NR_A(CH₂)_OOC(＝O)R_B、C(＝O)NR_A(CH₂)_OC(＝O)OR_B、C(＝O)NR_A(CH₂)_OC(＝O)NR_AR_B、C(＝O)NR_A(CH₂)_OOC(＝O)(CH₂)_OOR_BOr

R_HIndependently for each occurrence is halogen;

z' is independently CH at each occurrence₂、O、S、NR_A、N(CH₂)_oOR_A、N(CH₂)_oNR_AR_B、N(CH₂)_oCOOR_A、N(CH₂)_oOC(＝O)R_A、N(CH₂)_oCONR_AR_B、N(CH₂)_oNR_AC(＝O)R_BOr N (CH)₂)_oOC(＝O)(CH₂)_oOR_A；

o is independently at each occurrence 0,1, 2,3, 4, 5 or 6;

p is independently at each occurrence an integer of 0,1, 2,3, 4, or 5; and is

m is independently at each occurrence an integer of 1, 2,3, 4 or 5.

In certain embodiments, the compounds disclosed herein have the structure of formula (II) or (III):

or a pharmaceutically acceptable salt thereof, wherein:

x is 0 or 1;

y is H OR OR₅(ii) a Wherein R is₅Is H or methyl;

m 'and n' are each independently 0,1, 2,3, 4, 5,6, 7,8, 9, 10, 11, or 12;

R_A’and R_B’Independently for each occurrence:

hydrogen;

(C₁-C₆) Alkyl, optionally substituted by one or more radicals R which may be the same or different_DSubstitution;

(C₂-C₆) Alkenyl or (C)₂-C₆) An alkynyl group;

optionally is (C)₁-C₆) Alkyl substituted (C)₃-C₇) A cycloalkyl group;

optionally substituted by a group selected from halogen, -O (C)₁-C₆) Alkyl, -C (═ O) O (C)₁-C₆) Phenyl or benzyl of alkyl, amino, alkylamino and dialkylamino, which may be substituted identically or differently by 1 to 5 radicals;

or R_A’And R_B’Together with the nitrogen atom to which they are attached form a saturated or unsaturated heterocyclic ring containing from 3 to 7 ring atoms, which may optionally contain another heteroatom selected from nitrogen, oxygen and sulfur and may optionally be substituted by 1 to 4 groups, which may be the same or different, selected from alkyl, phenyl and benzyl; and is

R_DIndependently at each occurrence is halogen, hydroxy, O (C)₁-C₄) Alkyl, C (═ O) (C)₁-C₄) Alkyl, C (═ O) O (C)₁-C₄) Alkyl orWherein Z is CH₂、O、S、NH、NCH₃NEt, N-isopropyl, N-neopentyl, N-CH₂CH₂OH or N-CH₂CH₂OMe。

In certain embodiments, the compounds disclosed herein have the structure of formula (IV) or (V):

(ii) a Wherein x is 0 or 1; y is H OR OR₅(ii) a Wherein R is₅Is H or methyl; r_1’Each occurrence is independently H, halogen, aryl OR substituted aryl, heteroaryl OR substituted heteroaryl, OR_A、SR_A、NR_AR_B、-NR_A(CH₂)_OOR_B、-N((CH₂)_OOR_A)((CH₂)_OOR_B)-C(＝O)R_A、-C(＝O)OR_A、-OC(＝O)R_A、-OC(＝O)(C₁-C₆alkyl-R_H)、-OC(＝O)(CH₂)_oOR_A、-C(＝O)NR_AR_B、-NR_AC(＝O)R_B、-N(C(＝O)R_A)(C(＝O)R_B)、-N(C(＝O)(C₁-C₆alkyl-R_H))(C(＝O)(C₁-C₆alkyl-R_H))、-N(C(＝O)(C₁-C₆alkyl-R_H))₂、-NR_AC(＝O)(CH₂)_OOR_B、-NR_AC(＝O)(CH₂)_OOR_B、-N(C(＝O)(CH₂)_OOR_B)₂、-NR_AC(＝O)(CH₂)_ONR_AR_B、-NR_A(CH₂)_OC(＝O)OR_B、-N((CH₂)_OC(＝O)OR_A)((CH₂)_OC(＝O)OR_B)、-NR_A(CH₂)_OC(＝O)NR_AR_B、-N((CH₂)_OC(＝O)NR_AR_B)((CH₂)_OC(＝O)NR_AR_B)、-NR_A(CH₂)_OC(＝O)NR_A(CH₂)_OOR_B、-C(＝O)N((CH₂)_OOR_B)₂、-N((CH₂)_OC(＝O)NR_A(CH₂)_OOR_B)((CH₂)_OC(＝O)NR_A(CH₂)_OOR_B)、-C(＝O)NR_A(CH₂)_OOR_B、-C(＝O)NR_A(CH₂)_OOR_B、-C(＝O)NR_A(CH₂)_ONR_AR_B、-N＝CR_A-NR_AR_B、-NR_B-C(＝NH)-NR_AR_B、O(CH₂)_mOR_A、O(CH₂)_mCOOR_A、O(CH₂)_mCONR_AR_B、O(CH₂)_mCONR_A(CH₂)_mOR_A、O(CH₂)_mO(CH₂)_mOR_A、O(CH₂)_mNR_AR_B、O(CH₂)_mO(CH₂)_mNR_AR_B、NR_C(CH₂)_mNR_AR_B、NR_C(CH₂)_mNR_C(CH₂)_mNR_AR_B、Wherein said aryl or heteroaryl is optionally substituted by a substituent selected from the group consisting of halogen, hydroxy, (C)₁-C₆) Alkyl group, (CH)₂)_pOR_A、(CH₂)_pNR_AR_B、(CH₂)_pC(＝O)NR_AR_BAnd (CH)₂)_pC(＝O)OR_AMay be the same or different.

In certain embodiments, R₁is-NR_AC(＝O)R_BAnd R is_1’Is OR_A。R_ANon-limiting examples of (d) include H, Me, Et, Pr, Bu, and pentyl. R_BNon-limiting examples of (d) include H, Me, Et, Pr, Bu, and pentyl. In certain embodiments, wherein R is₁is-NHC (═ O) R_BAnd R is_1’Is OH; and Y is OH or H.

In certain embodiments, the compounds disclosed herein have the structure of formula (VI):

wherein

x is 0 or 1;

w is CH₂O or S;

y is H OR OR₅(ii) a Wherein R is₅Is H or methyl;

m 'and n' are each independently 0,1, 2,3, 4, 5,6, 7,8, 9, or 10; and is

R₁Each occurrence is independently H, halogen, aryl OR substituted aryl, heteroaryl OR substituted heteroaryl, OR_A、SR_A、NR_AR_B、-C(＝O)R_A、-C(＝O)OR_A、-C(＝O)NR_AR_B、-NR_AC(＝O)R_B、-NR_AC(＝O)(CH₂)_OR_B、-NR_AC(＝O)(CH₂)_OOR_B、-NR_AC(＝O)(CH₂)_ONR_AR_Bor-C (═ O) NR_A(CH₂)_OR_B。

In another aspect, the invention provides a pharmaceutical composition comprising at least one compound as described herein and a pharmaceutically acceptable carrier.

In yet another aspect, the present invention provides a method of treating or preventing a viral infection in a mammalian species in need thereof, the method comprising administering to the mammalian species a therapeutically effective amount of at least one compound as described herein.

In another aspect, the present invention provides a method of treating or preventing hepatitis c virus infection in a mammalian species in need thereof, the method comprising administering to the mammalian species a therapeutically effective amount of at least one compound as described herein.

In another aspect, the present invention provides a method of inhibiting cyclophilin in a subject in need thereof, comprising administering to the subject an amount of at least one compound as described herein effective to inhibit cyclophilin.

In another aspect, the present invention provides a method of treating or preventing a cyclophilin-mediated disease in a mammalian species in need thereof, comprising administering to the mammalian species a therapeutically effective amount of at least one compound as described herein.

In another aspect, the present invention provides a method of treating or preventing a disease in a mammalian species in need thereof, the method comprising administering to the mammalian species a therapeutically effective amount of at least one compound as described herein, wherein the disease is selected from the group consisting of inflammation, respiratory inflammation, rheumatoid arthritis, and dry eye.

In yet another aspect, the invention provides a method of treating or preventing a disease in a mammalian species in need thereof, the method comprising administering to the mammalian species a therapeutically effective amount of at least one compound as described herein, wherein the disease is selected from the group consisting of neurodegenerative diseases, such as alzheimer's disease, parkinson's disease, huntington's disease, and ALS; traumatic brain injury; stroke; and ischemia reperfusion injury in the brain, heart and kidney.

In a further aspect, the present invention provides a method of treating or preventing a disease in a mammalian species in need thereof, the method comprising administering to said mammalian species a therapeutically effective amount of at least one compound as described herein, wherein said disease is selected from cardiovascular disease, vascular stenosis, atherosclerosis, abdominal aortic aneurysm, cardiac hypertrophy, aortic rupture, pulmonary hypertension, myocarditis and myocardial fibrosis, and ischemic heart disease.

In yet another aspect, the invention provides a method of treating or preventing a disease or condition in a mammalian species in need thereof, the method comprising administering to the mammalian species a therapeutically effective amount of at least one compound as described herein, wherein the disease or condition is selected from cancer; obesity; diabetes mellitus; muscular dystrophy; lung and liver and kidney diseases and their protection; and alopecia.

In yet another aspect, the invention provides a method of treating or preventing a disease or condition in a mammalian species in need thereof, the method comprising administering to the mammalian species a therapeutically effective amount of at least one compound as described herein, wherein the disease or condition is selected from allergic conjunctivitis, atopic and vernal keratoconjunctivitis, atopic keratoconjunctivitis, anterior uveitis, behcet's disease, blepharitis, chronic ocular surface inflammation resulting from viral infection, corneal graft rejection, impaired corneal sensitivity due to surgery of the cornea or other ocular surface, meibomian gland disease, pterygium (ptyregia), ocular symptoms of graft versus host disease, ocular allergy, ocular cicatricial pemphigoid, Steven's syndrome, vernal keratoconjunctivitis, uveitis, herpes simplex keratitis, ocular rosacea, and meibomian spots.

Detailed Description

Definition of

The following are definitions of terms used in this specification. Unless otherwise indicated, the initial definitions of a group or term provided herein apply to that group or term alone or as part of another group throughout this specification.

The terms "alkyl" and "alkane" refer to straight or branched chain alk (hydrocarbon) yl groups containing 1 to 12 carbon atoms, preferably 1 to 6 carbon atoms. Exemplary "alkyl" groups include methyl, ethyl, propyl, isopropyl, n-butyl, t-butyl, isobutylpentyl, hexyl, isohexyl, heptyl, 4, 4-dimethylpentyl, octyl, 2,2, 4-trimethylpentyl, nonyl, decyl, undecyl, dodecyl and the like. Term "(C)₁-C₄) Alkyl "means a straight or branched chain alk (en) yl group containing 1 to 4 carbon atoms, such as methyl, ethyl, propyl, isopropyl, n-butyl, t-butyl and isobutyl. Term "(C)₁-C₆) Alkyl "means a straight or branched chain alk (en) yl group containing 1 to 6 carbon atoms, except for being" (C)₁-C₄) Alkyl groups "are other than those exemplified, for example n-hexyl, 2-methylpentyl, 3-methylpentyl, 4-methylpentyl, 2, 3-dimethylbutyl, 2, 2-dimethylbutyl. "substituted alkyl" refers to an alkyl group substituted with one or more substituents, preferably 1 to 4 substituents, at any available point of attachment. Exemplary substituents include, but are not limited to, one or more of the following groups: hydrogen, halogen (e.g. single halogen substituent or multiple halogen substituents, in the latter case forming, for example, CF)₃Or with Cl₃Alkyl group of (i), cyano, nitro, oxo (i.e., ═ O), CF₃，OCF₃Cycloalkyl, alkenyl, cycloalkenyl, alkynyl, heterocycle, aryl, OR_a，SR_a，S(＝O)R_e，S(＝O)₂R_e，P(＝O)₂R_e，S(＝O)₂OR_e，P(＝O)₂OR_e，NR_bR_c，NR_bS(＝O)₂R_e，NR_bP(＝O)₂R_e，S(＝O)₂NR_bR_c，P(＝O)₂NR_bR_c，C(＝O)OR_d，C(＝O)R_a，C(＝O)NR_bR_c，OC(＝O)R_a，OC(＝O)NR_bR_c，NR_bC(＝O)OR_e，NR_dC(＝O)NR_bR_c，NR_dS(＝O)₂NR_bR_c，NR_dP(＝O)₂NR_bR_c，NR_bC(＝O)R_aOr NR_bP(＝O)₂R_eWherein R is_aEach occurrence of (a) is independently hydrogen, alkyl, cycloalkyl, alkenyl, cycloalkenyl, alkynyl, heterocycle or aryl; r_b，R_cAnd R_dEach occurrence of (A) is independently hydrogen, alkyl, cycloalkyl, heterocycle, aryl, or said R_bAnd R_cOptionally forming a heterocyclic ring together with the N to which they are bonded; and R is_eIndependently for each occurrence of (A) is alkyl, cycloalkyl, alkenylCycloalkenyl, alkynyl, heterocycle or aryl. Among the above exemplary substituents, groups such as alkyl, cycloalkyl, alkenyl, alkynyl, cycloalkenyl, heterocycle and aryl may themselves be optionally substituted.

The term "alkenyl" refers to a straight or branched chain hydrocarbon group containing 2 to 12 carbon atoms and at least one carbon-carbon double bond. Examples of such groups include vinyl or allyl. The term "C₂-C₆Alkenyl "means a straight or branched chain hydrocarbon group containing 2 to 6 carbon atoms and at least one carbon-carbon double bond, such as ethylene, propenyl, 2-propenyl, (E) -but-2-enyl, (Z) -but-2-enyl, 2-methyl (E) -but-2-enyl, 2-methyl (Z) -but-2-enyl, 2, 3-dimethyl-but-2-enyl, (Z) -pent-2-enyl, (E) -pent-1-enyl, (Z) -hex-1-enyl, (E) -pent-2-enyl, (Z) -hex-2-enyl, (E) -hex-2-enyl, (Z) -hex-1-enyl, (E) -hex-1-enyl, (Z) -hex-3-enyl, (E) -hex-3-enyl and (E) -hex-1, 3-dienyl. "substituted alkenyl" refers to alkenyl substituted with one or more substituents at any available point of attachment, preferably 1 to 4 substituents. Exemplary substituents include, but are not limited to, one or more of the following groups: hydrogen, halogen (e.g. single halogen substituent or multiple halogen substituents, in the latter case forming, for example, CF)₃Or with Cl₃Alkyl group of (i), cyano, nitro, oxo (i.e., ═ O), CF₃，OCF₃Cycloalkyl, alkenyl, cycloalkenyl, alkynyl, heterocycle, aryl, OR_a，SR_a，S(＝O)R_e，S(＝O)₂R_e，P(＝O)₂R_e，S(＝O)₂OR_e，P(＝O)₂OR_e，NR_bR_c，NR_bS(＝O)₂R_e，NR_bP(＝O)₂R_e，S(＝O)₂NR_bR_c，P(＝O)₂NR_bR_c，C(＝O)OR_d，C(＝O)R_a，C(＝O)NR_bR_c，OC(＝O)R_a，OC(＝O)NR_bR_c，NR_bC(＝O)OR_e，NR_dC(＝O)NR_bR_c，NR_dS(＝O)₂NR_bR_c，NR_dP(＝O)₂NR_bR_c，NR_bC(＝O)R_aOr NR_bP(＝O)₂R_eWherein R is_aEach occurrence of (a) is independently hydrogen, alkyl, cycloalkyl, alkenyl, cycloalkenyl, alkynyl, heterocycle or aryl; r_b，R_cAnd R_dEach occurrence of (A) is independently hydrogen, alkyl, cycloalkyl, heterocycle, aryl, or said R_bAnd R_cOptionally forming a heterocyclic ring together with the N to which they are bonded; and R is_eEach occurrence of (a) is independently alkyl, cycloalkyl, alkenyl, cycloalkenyl, alkynyl, heterocycle, or aryl. The above exemplary substituents may themselves be optionally substituted.

The term "alkynyl" refers to a straight or branched chain hydrocarbon radical containing from 2 to 12 carbon atoms and at least one carbon-carbon triple bond. Examples of such groups include ethynyl. The term "C₂-C₆Alkynyl "means a straight or branched chain hydrocarbon group containing from 2 to 6 carbon atoms and at least one carbon-carbon triple bond, such as ethynyl, prop-1-ynyl, prop-2-ynyl, but-1-ynyl, but-2-ynyl, pent-1-ynyl, pent-2-ynyl, hex-1-ynyl, hex-2-ynyl, hex-3-ynyl. "substituted alkynyl" refers to alkynyl substituted with one or more substituents at any available point of attachment, preferably 1 to 4 substituents. Exemplary substituents include, but are not limited to, one or more of the following groups: hydrogen, halogen (e.g. single halogen substituent or multiple halogen substituents, in the latter case forming, for example, CF)₃Or with Cl₃Alkyl group of (i), cyano, nitro, oxo (i.e., ═ O), CF₃，OCF₃Cycloalkyl, alkenyl, cycloalkenyl, alkynyl, heterocycle, aryl, OR_a，SR_a，S(＝O)R_e，S(＝O)₂R_e，P(＝O)₂R_e，S(＝O)₂OR_e，P(＝O)₂OR_e，NR_bR_c，NR_bS(＝O)₂R_e，NR_bP(＝O)₂R_e，S(＝O)₂NR_bR_c，P(＝O)₂NR_bR_c，C(＝O)OR_d，C(＝O)R_a，C(＝O)NR_bR_c，OC(＝O)R_a，OC(＝O)NR_bR_c，NR_bC(＝O)OR_e，NR_dC(＝O)NR_bR_c，NR_dS(＝O)₂NR_bR_c，NR_dP(＝O)₂NR_bR_c，NR_bC(＝O)R_aOr NR_bP(＝O)₂R_eWherein R is_aEach occurrence of (a) is independently hydrogen, alkyl, cycloalkyl, alkenyl, cycloalkenyl, alkynyl, heterocycle or aryl; r_b，R_cAnd R_dEach occurrence of (A) is independently hydrogen, alkyl, cycloalkyl, heterocycle, aryl, or said R_bAnd R_cOptionally forming a heterocyclic ring together with the N to which they are bonded; and R is_eEach occurrence of (a) is independently alkyl, cycloalkyl, alkenyl, cycloalkenyl, alkynyl, heterocycle, or aryl. The above exemplary substituents may themselves be optionally substituted.

The term "cycloalkyl" refers to a fully saturated cyclic hydrocarbon group containing from 1 to 4 rings and from 3 to 8 carbon atoms per ring. "C₃-C₇Cycloalkyl "means cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl or cycloheptyl. "substituted cycloalkyl" refers to a cycloalkyl group substituted at any available point of attachment with one or more substituents, preferably 1 to 4 substituents. Exemplary substituents include, but are not limited to, one or more of the following groups: hydrogen, halogen (e.g. single halogen substituent or multiple halogen substituents, in the latter case forming, for example, CF)₃Or with Cl₃Alkyl group of (i), cyano, nitro, oxo (i.e., ═ O), CF₃，OCF₃Cycloalkyl, alkenyl, cycloalkenyl, alkynyl, heterocycle, aryl, OR_a，SR_a，S(＝O)R_e，S(＝O)₂R_e，P(＝O)₂R_e，S(＝O)₂OR_e，P(＝O)₂OR_e，NR_bR_c，NR_bS(＝O)₂R_e，NR_bP(＝O)₂R_e，S(＝O)₂NR_bR_c，P(＝O)₂NR_bR_c，C(＝O)OR_d，C(＝O)R_a，C(＝O)NR_bR_c，OC(＝O)R_a，OC(＝O)NR_bR_c，NR_bC(＝O)OR_e，NR_dC(＝O)NR_bR_c，NR_dS(＝O)₂NR_bR_c，NR_dP(＝O)₂NR_bR_c，NR_bC(＝O)R_aOr NR_bP(＝O)₂R_eWherein R is_aEach occurrence of (a) is independently hydrogen, alkyl, cycloalkyl, alkenyl, cycloalkenyl, alkynyl, heterocycle or aryl; r_b，R_cAnd R_dEach occurrence of (A) is independently hydrogen, alkyl, cycloalkyl, heterocycle, aryl, or said R_bAnd R_cOptionally forming a heterocyclic ring together with the N to which they are bonded; and R is_eEach occurrence of (a) is independently alkyl, cycloalkyl, alkenyl, cycloalkenyl, alkynyl, heterocycle, or aryl. The above exemplary substituents may themselves be optionally substituted. Exemplary substituents also include spiro-linked or fused cyclic substituents, particularly spiro-linked cycloalkyl, spiro-linked cycloalkenyl, spiro-linked heterocycle (without heteroaryl), fused cycloalkyl, fused cycloalkenyl, fused heterocycle, or fused aryl, wherein the above cycloalkyl, cycloalkenyl, heterocycle, and aryl substituents themselves may be optionally substituted.

The term "cycloalkenyl" refers to partially unsaturated cyclic hydrocarbon groups containing from 1 to 4 rings and from 3 to 8 carbon atoms per ring. Examples of such groups include cyclobutenyl, cyclopentenyl, cyclohexenyl, and the like. "substituted cycloalkenyl" refers to cycloalkenyl substituted with one or more substituents at any available point of attachment, preferably 1 to 4 substituents. Exemplary substituents include, but are not limited to, one or more of the following groups: hydrogen, halogen (e.g. single halogen substituent or multiple halogen substituents, in the latter case forming, for example, CF)₃Or with Cl₃Alkyl group of (i), cyano, nitro, oxo (i.e., ═ O), CF₃，OCF₃Cycloalkyl, alkenyl, cycloalkenyl, alkynyl, heterocycle, aryl, OR_a，SR_a，S(＝O)R_e，S(＝O)₂R_e，P(＝O)₂R_e，S(＝O)₂OR_e，P(＝O)₂OR_e，NR_bR_c，NR_bS(＝O)₂R_e，NR_bP(＝O)₂R_e，S(＝O)₂NR_bR_c，P(＝O)₂NR_bR_c，C(＝O)OR_d，C(＝O)R_a，C(＝O)NR_bR_c，OC(＝O)R_a，OC(＝O)NR_bR_c，NR_bC(＝O)OR_e，NR_dC(＝O)NR_bR_c，NR_dS(＝O)₂NR_bR_c，NR_dP(＝O)₂NR_bR_c，NR_bC(＝O)R_aOr NR_bP(＝O)₂R_eWherein R is_aEach occurrence of (a) is independently hydrogen, alkyl, cycloalkyl, alkenyl, cycloalkenyl, alkynyl, heterocycle or aryl; r_b，R_cAnd R_dEach occurrence of (A) is independently hydrogen, alkyl, cycloalkyl, heterocycle, aryl, or said R_bAnd R_cOptionally forming a heterocyclic ring together with the N to which they are bonded; and R is_eEach occurrence of (a) is independently alkyl, cycloalkyl, alkenyl, cycloalkenyl, alkynyl, heterocycle, or aryl. The above exemplary substituents may themselves be optionally substituted. Exemplary substituents also include spiro-linked or fused cyclic substituents, particularly spiro-linked cycloalkyl, spiro-linked cycloalkenyl, spiro-linked heterocycle (without heteroaryl), fused cycloalkyl, fused cycloalkenyl, fused heterocycle, or fused aryl, wherein the above cycloalkyl, cycloalkenyl, heterocycle, and aryl substituents themselves may be optionally substituted.

The term "aryl" refers to a cyclic aromatic hydrocarbon group having 1 to 5 aromatic rings, especially a monocyclic or bicyclic group, such as phenyl, biphenyl or naphthyl. When containing two or more aromatic rings (bicyclic, etc.), theThe aromatic rings of the aryl groups can be joined at a single point (e.g., biphenyl), or fused (e.g., naphthyl, phenanthrenyl, etc.). "substituted aryl" refers to an aryl group substituted with one or more substituents, preferably 1 to 3 substituents, at any available point of attachment. Exemplary substituents include, but are not limited to, one or more of the following groups: hydrogen, halogen (e.g. single halogen substituent or multiple halogen substituents, in the latter case forming, for example, CF)₃Or with Cl₃Alkyl group of (i), cyano, nitro, oxo (i.e., ═ O), CF₃，OCF₃Cycloalkyl, alkenyl, cycloalkenyl, alkynyl, heterocycle, aryl, OR_a，SR_a，S(＝O)R_e，S(＝O)₂R_e，P(＝O)₂R_e，S(＝O)₂OR_e，P(＝O)₂OR_e，NR_bR_c，NR_bS(＝O)₂R_e，NR_bP(＝O)₂R_e，S(＝O)₂NR_bR_c，P(＝O)₂NR_bR_c，C(＝O)OR_d，C(＝O)R_a，C(＝O)NR_bR_c，OC(＝O)R_a，OC(＝O)NR_bR_c，NR_bC(＝O)OR_e，NR_dC(＝O)NR_bR_c，NR_dS(＝O)₂NR_bR_c，NR_dP(＝O)₂NR_bR_c，NR_bC(＝O)R_aOr NR_bP(＝O)₂R_eWherein R is_aEach occurrence of (a) is independently hydrogen, alkyl, cycloalkyl, alkenyl, cycloalkenyl, alkynyl, heterocycle or aryl; r_b，R_cAnd R_dEach occurrence of (A) is independently hydrogen, alkyl, cycloalkyl, heterocycle, aryl, or said R_bAnd R_cOptionally forming a heterocyclic ring together with the N to which they are bonded; and R is_eEach occurrence of (a) is independently alkyl, cycloalkyl, alkenyl, cycloalkenyl, alkynyl, heterocycle, or aryl. The above exemplary substituents may themselves be optionally substituted. Exemplary substituents also include fused cyclic groups, especially fused cycloalkyl, fusedCycloalkenyl, fused heterocycle or fused aryl, wherein the above cycloalkyl, cycloalkenyl, heterocycle and aryl substituents themselves may be optionally substituted.

The terms "heterocycle" and "heterocyclic" refer to a fully saturated or partially or fully unsaturated, including aromatic (i.e., "heteroaryl"), cyclic group (e.g., a 4-to 7-membered monocyclic, 7-to 11-membered bicyclic, or 8-to 16-membered tricyclic ring system) having at least one heteroatom in at least one carbon atom-containing ring. Each ring of the heterocyclic group containing a heteroatom may have 1, 2,3 or 4 heteroatoms selected from nitrogen atoms, oxygen atoms and/or sulfur atoms, where the nitrogen and sulfur heteroatoms may optionally be oxidized and the nitrogen heteroatoms may optionally be quaternized. (the term "heteroarylcation" refers to a heteroaryl group that carries a quaternary nitrogen atom and thus a positive charge). The heterocyclic group may be attached to the rest of the molecule at any heteroatom or carbon atom of the ring or ring system. Exemplary monocyclic heterocyclic groups include azetidinyl (azetidinyl), pyrrolidinyl, pyrrolyl, pyrazolyl, oxetanyl (oxyethanyl), pyrazolinyl, imidazolyl, imidazolinyl, imidazolidinyl, oxazolyl, oxazolidinyl, isoxazolidinyl, isoxazolyl, thiazolyl, thiadiazolyl (thiadiazolyl), tetrahydrothiazolyl, isothiazolyl, furyl, tetrahydrofuryl, thienyl, oxadiazolyl (oxadiazolyl), piperidinyl, piperazinyl, 2-oxopiperazinyl, 2-oxopiperidinyl, 2-oxopyrrolidinyl (2-oxopyrodinyl), 2-oxoazepinyl, aza (azepinyl), hexahydrodiazepinyl, 4-piperidonyl (4-piperidonyl), pyridinyl, pyrazinyl, pyrimidinyl, pyridazinyl, triazinyl, triazolyl, tetrazolyl, tetrahydropyranyl, morpholinyl, thiomorpholinyl (thiomorpholinyl), thiomorpholinyl sulfoxide, thiomorpholinyl sulfone, 1, 3-dioxolane, and tetrahydro-1, 1-dioxothienyl, and the like. Exemplary bicyclic heterocyclic groups include indolyl, isoindolyl, benzothiazolyl, benzoxazolyl, benzoxadiazolyl, benzothienyl, benzo [ d ] [1,3] dioxolanyl (dioxolyl), 2, 3-dihydrobenzo [ b ] [1,4] dioxinyl (dioxinyl), quinuclidinyl, quinolinyl, tetrahydroisoquinolinyl (tetrahydroquinonyl), isoquinolinyl (isoquinonyl), benzimidazolyl, benzopyranyl, indolizinyl, benzofuranyl, benzofuropinyl, chromonyl (chromanyl), coumarinyl (chromanyl), benzopyranyl, cinnolinyl, quinoxalinyl, indazolyl, pyrrolopyridinyl, furopyridinyl (such as furo [2,3-c ] pyridinyl, furo [3,2-b ] pyridinyl ] or furo [2,3-b ] pyridinyl), dihydroisoindolyl, dihydroquinazolinyl (e.g., 3, 4-dihydro-4-oxo-quinazolinyl), triazinyl azepinyl (triazinylazepinyl), tetrahydroquinolinyl (tetrahydroquinolinyl), and the like. Exemplary tricyclic heterocyclic groups include carbazolyl, benzidolyl, phenanthryl, acridinyl, phenanthridinyl, xanthenyl and the like.

"substituted heterocycle" and "substituted heterocycle" (e.g., "substituted heteroaryl") refer to a heterocycle or heterocyclic group substituted at any available point of attachment with one or more substituents, preferably 1 to 4 substituents. Exemplary substituents include, but are not limited to, one or more of the following groups: hydrogen, halogen (e.g. single halogen substituent or multiple halogen substituents, in the latter case forming, for example, CF)₃Or with Cl₃Alkyl group of (i), cyano, nitro, oxo (i.e., ═ O), CF₃，OCF₃Cycloalkyl, alkenyl, cycloalkenyl, alkynyl, heterocycle, aryl, OR_a，SR_a，S(＝O)R_e，S(＝O)₂R_e，P(＝O)₂R_e，S(＝O)₂OR_e，P(＝O)₂OR_e，NR_bR_c，NR_bS(＝O)₂R_e，NR_bP(＝O)₂R_e，S(＝O)₂NR_bR_c，P(＝O)₂NR_bR_c，C(＝O)OR_d，C(＝O)R_a，C(＝O)NR_bR_c，OC(＝O)R_a，OC(＝O)NR_bR_c，NR_bC(＝O)OR_e，NR_dC(＝O)NR_bR_c，NR_dS(＝O)₂NR_bR_c，NR_dP(＝O)₂NR_bR_c，NR_bC(＝O)R_aOr NR_bP(＝O)₂R_eWherein R is_aEach occurrence of (a) is independently hydrogen, alkyl, cycloalkyl, alkenyl, cycloalkenyl, alkynyl, heterocycle or aryl; r_b，R_cAnd R_dEach occurrence of (A) is independently hydrogen, alkyl, cycloalkyl, heterocycle, aryl, or said R_bAnd R_cOptionally forming a heterocyclic ring together with the N to which they are bonded; and R is_eEach occurrence of (a) is independently alkyl, cycloalkyl, alkenyl, cycloalkenyl, alkynyl, heterocycle, or aryl. Exemplary substituents may themselves be optionally substituted. Exemplary substituents also include spiro-linked or fused cyclic substituents at any available point(s) of attachment, particularly spiro-linked cycloalkyl, spiro-linked cycloalkenyl, spiro-linked heterocycle (without heteroaryl), fused cycloalkyl, fused cycloalkenyl, fused heterocycle, or fused aryl, where the above cycloalkyl, cycloalkenyl, heterocycle and aryl substituents themselves may be optionally substituted.

The term "alkylamino" refers to a group having the structure-NHR ', wherein R' is hydrogen, alkyl or substituted alkyl, cycloalkyl or substituted cycloalkyl, as defined herein. Examples of alkylamino include, but are not limited to, methylamino, ethylamino, n-propylamino, isopropylamino, cyclopropylamino, n-butylamino, tert-butylamino, neopentylamino, n-pentylamino, hexylamino, cyclohexylamino, and the like.

The term "dialkylamino" refers to a group having the structure-NRR ', wherein R and R' are each independently alkyl or substituted alkyl, cycloalkyl or substituted cycloalkyl, cycloalkenyl or substituted cycloalkenyl, aryl or substituted aryl, heterocyclyl or substituted heterocyclyl, as defined herein. R and R' may be the same or different in the dialkylamino moiety. Examples of dialkylamino include, but are not limited to, dimethylamino, methylethylamino, diethylamino, methylpropylamino, di (n-propyl) amino, di (isopropyl) amino, di (cyclopropyl) amino, di (n-butyl) amino, di (tert-butyl) amino, di (neopentyl) amino, di (n-pentyl) amino, di (hexyl) amino, di (cyclohexyl) amino, and the like. In certain embodiments, R and R' are joined to form a cyclic structure. The resulting cyclic structure may be aromatic or non-aromatic. Examples of cyclic diaminoalkyl groups include, but are not limited to, aziridinyl, pyrrolidinyl, piperidinyl, morpholinyl, pyrrolyl, imidazolyl, 1,3, 4-triazolyl (1,3,4-trianolyl), and tetrazolyl.

The term "halogen" or "halo" refers to chlorine, bromine, fluorine or iodine.

Unless otherwise indicated, it is assumed that any heteroatom having an unsaturated valence has a hydrogen atom sufficient to satisfy the valence.

The compounds of the present invention may form salts, which are also within the scope of the present invention. Unless otherwise indicated, reference to a compound of the invention is understood to refer to a salt thereof. The term "salt(s)", as used herein, denotes acidic and/or basic salts formed with inorganic and/or organic acids and bases. Furthermore, when a compound of the present invention comprises both a basic moiety, such as, but not limited to, a pyridine or imidazole, and an acidic moiety, such as, but not limited to, a carboxylic acid, zwitterions ("inner salts") may be formed and are included within the term(s) "salt(s)" as used herein. Pharmaceutically acceptable (i.e., non-toxic, physiologically acceptable) salts are preferred, although other salts are also useful, for example, in isolation or purification steps that may be employed in the preparation process. Salts of the compounds of the invention may be formed, for example, by reacting a compound of formula I with an amount (e.g., equivalent amount) of an acid or base, for example, in a medium in which the salt precipitates or in an aqueous medium which is subsequently lyophilized.

Compounds of the invention containing a basic moiety such as, but not limited to, an amine or a pyridine or imidazole ring can form salts with various organic and inorganic acids. Exemplary acid addition salts include acetates (such as those formed with acetic acid or trihaloacetic acids, e.g., trifluoroacetic acid), adipates, alginates, ascorbates, aspartates, benzoates, benzenesulfonates, bisulfates, borates, butyrates, citrates, camphorates (camphostates), camphorsulfonates, cyclopentanepropionates, digluconates, dodecylsulfates, ethanesulfonates, fumarates, glucoheptonates (glucoheptanates), glycerophosphates, hemisulfates (hemisulfates), heptanoates (heptanates), hexanoates, hydrochlorides, hydrobromides, hydroiodides, hydroxyethanesulfonates (e.g., 2-hydroxyethanesulfonates), lactates, maleates, methanesulfonates, naphthalenesulfonates (e.g., 2-naphthalenesulfonates), nicotinates, nitrates, oxalates, pectinates (pectinates), persulfates, phenylpropionates (e.g., 3-phenylpropionate), phosphates, picrates, pivalates, propionates, salicylates, succinates, sulfates (such as those formed with sulfuric acid), sulfonates, tartrates, thiocyanates, tosylates, undecanoates, and the like.

The compounds of the invention containing an acidic moiety, such as, but not limited to, a carboxylic acid, can form salts with a variety of organic and inorganic bases. Exemplary basic salts include ammonium salts, alkali metal salts such as sodium, lithium and potassium salts, alkaline earth metal salts such as calcium and magnesium salts, salts with organic bases (e.g., organic amines) such as benzathine, dicyclohexylamine, hydrabamines (formed with N, N-bis (dehydroabietyl) ethylenediamine), N-methyl-D-glucamine, N-methyl-D-imidazole bisamides (glycamides), t-butylamine, and salts with amino acids such as arginine, lysine, and the like. Basic nitrogen-containing groups can be quaternized with agents such as lower alkyl halides (e.g., methyl, ethyl, propyl, and butyl chlorides, bromides, and iodides), dialkyl sulfates (e.g., dimethyl, diethyl, dibutyl, and diamyl sulfates), long chain halides (e.g., decyl, lauryl, myristyl and stearyl chlorides, bromides and iodides), aralkyl halides (e.g., benzyl and phenethyl bromides), and others.

Prodrugs and solvates of the compounds of the invention are also contemplated herein. The term "prodrug" as used herein denotes a compound that undergoes a chemical transformation by metabolic or chemical processes to yield a compound of the invention, or a salt and/or solvate thereof, upon administration to a subject. Solvates of the compounds of the invention include, for example, hydrates.

The compounds of the invention, and salts or solvates thereof, may exist in their tautomeric form (for example as an amide or imino ether). All such tautomeric forms are contemplated herein as part of the present invention.

All stereoisomers of the compounds of the present invention (e.g., those that may exist due to asymmetric carbon atoms on various substituents), including enantiomeric and diastereomeric forms, are contemplated within the scope of the invention. Individual stereoisomers of the compounds of the invention may, for example, be substantially free of other isomers (e.g., as pure or substantially pure optical isomers with a particular activity), or may be mixed, for example, as racemates or with all other or other selected stereoisomers. The chiral centers of the present invention may have the S or R configuration as defined by the International Union of Pure and Applied Chemistry (IUPAC)1974 recommendation. The racemic forms can be resolved by physical methods such as, for example, fractional crystallization, separation or crystallization of diastereomeric derivatives, or separation by chiral column chromatography. The individual optical isomers may be obtained from the racemates by any suitable method, including, but not limited to, conventional methods such as, for example, salt formation with an optically active acid followed by crystallization.

Preferably, after preparing the compounds of the present invention, they are isolated and purified to give a composition containing an amount of equal to or greater than 90%, e.g., equal to or greater than 95%, equal to or greater than 99% pure by weight ("substantially pure" compound I), which is then used or formulated as described herein. Such "substantially pure" compounds of the invention are also contemplated herein as part of the invention.

All configurational isomers of the compounds of the invention are contemplated, either in a mixture or in pure or substantially pure form. The definition of the compounds of the present invention encompasses cis (Z) and trans (E) alkene isomers, as well as cis and trans isomers of cyclic hydrocarbons or heterocycles.

Throughout the specification, their groups and substituents may be selected to provide stable moieties and compounds.

The definitions of specific functional groups and chemical terms are described in more detail below. For the purposes of the present invention, chemical elements are identified according to the CAS version of the periodic Table of elements in the 75 th edition of the handbook of chemistry and Physics, and specific functional groups are generally defined as described therein. In "Organic Chemistry", Thomas Sorrell, University Science Books, Sausaltito: the general principles of organic chemistry, as well as specific functional moieties and reactivity, are described in 1999, which is incorporated herein by reference in its entirety.

Certain compounds of the present invention may exist in specific geometric or stereoisomeric forms. The present invention contemplates all such compounds, including cis and trans isomers, R and S enantiomers, diastereomers, (D) -isomers, (L) -isomers, racemic mixtures thereof, and other mixtures thereof, as falling within the scope of the present invention. Additional asymmetric carbon atoms may be present in substituents such as alkyl groups. All such isomers and mixtures thereof are intended to be included in the present invention.

Mixtures of isomers containing any of a variety of isomer ratios may be used in accordance with the present invention. For example, when only two isomers are combined, mixtures containing ratios of 50:50, 60:40, 70:30, 80:20, 90:10, 95:5, 96:4, 97:3, 98:2, 99:1, or 100:0 isomers are contemplated by the present invention. Those of ordinary skill in the art will readily appreciate that similar ratios are contemplated for more complex isomer mixtures.

The invention also includes isotopically-labeled compounds, which are identical to those disclosed herein, but for the fact that one or more atoms are replaced by an atom having an atomic mass or mass number different from the atomic mass or mass number usually found in nature. Examples of isotopes that can be incorporated into the compounds of the invention include isotopes of hydrogen, carbon, nitrogen, oxygen, phosphorus, sulfur, fluorine and chlorine, respectivelySuch as²H、³H、¹³C、¹¹C、¹⁴C、¹⁵N、¹⁸O、¹⁷O、³¹P、³²P、³⁵S、¹⁸F and³⁶and (4) Cl. Compounds of the present invention containing the aforementioned isotopes and/or other isotopes of other atoms, or their enantiomers, diastereomers, tautomers or pharmaceutically acceptable salts or solvates, are within the scope of the present invention. Certain isotopically-labeled compounds of the present invention, for example, those into which a radioactive isotope such as³H and¹⁴c, useful in drug and/or substrate tissue distribution assays. Tritium (i.e. tritium³H) And carbon-14 (i.e.¹⁴C) Isotopes are particularly preferred for their ease of preparation and detection. In addition, heavier isotopes such as deuterium (i.e., deuterium) are used²H) Substitution may afford certain therapeutic advantages resulting from greater metabolic stability, such as increased in vivo half-life or reduced dosage requirements, and may therefore be preferred in certain circumstances. Isotopically labeled compounds can generally be prepared by carrying out the procedures disclosed in the schemes and/or in the examples below by substituting a readily available isotopically labeled reagent for a non-isotopically labeled reagent.

If, for example, a particular enantiomer of a compound of the invention is desired, it may be prepared by asymmetric synthesis or by derivatization with a chiral auxiliary, wherein the resulting diastereomeric mixture is separated and the auxiliary group is cleaved to provide the pure desired enantiomer. Alternatively, where the molecule contains a basic functionality, such as an amino group, or an acidic functionality, such as a carboxyl group, diastereomeric salts are formed with an appropriate optically active acid or base, followed by resolution of the diastereomers, whereby the pure enantiomers are formed by fractional crystallization or chromatographic means well known in the art, and subsequently recovered.

It is to be understood that a compound, as described herein, may be substituted with any number of substituents or functional moieties. In general, the term "substituted", whether preceded by the term "optionally", and including substituents in the general formulae of the present invention, means that the hydrogen group in a given structure is replaced by the group of the indicated substituent. When more than one position in any given structure may be substituted with more than one substituent selected from the group specified, the substituents at each position may be the same or different. As used herein, the term "substituted" is contemplated to include all permissible substituents of organic compounds. In a broad aspect, the permissible substituents include acyclic and cyclic, branched and unbranched, carbocyclic and heterocyclic, aromatic and nonaromatic substituents of organic compounds. For the purposes of the present invention, a heteroatom such as nitrogen may have a hydrogen substituent and/or any permissible substituents of organic compounds described herein that satisfy the valency of the heteroatom. Furthermore, the present invention is not intended to be limited in any way by the permissible substituents of organic compounds. Combinations of substituents and variables contemplated by the present invention are preferably those that result in the formation of stable compounds useful, for example, in the treatment of infectious diseases or proliferative disorders. The term "stable" as used herein preferably means that the compound has a stability for a period of time sufficient to allow manufacture and maintain the integrity of the compound long enough to be detected and preferably useful for the purposes described herein.

Compound (I)

The novel cyclosporin derivatives of the present invention are potent inhibitors of cyclophilins and are useful in the inhibition of viruses such as HCV, HBV and HIV.

In one aspect, the invention provides compounds of formula (I):

or a pharmaceutically acceptable salt thereof, wherein:

x is 0 or 1;

R₈is alkyl, alkenyl, alkynyl, cycloalkyl, cycloalkenyl, aryl or heteroaryl; wherein R is₈By one or more R₁Substitution; conditionIs R₈-R₁Is not n-butyl or (E) -but-2-enyl;

R₂is ethyl, 1-hydroxyethyl, isopropyl or n-propyl;

w is O, S or CH₂；

R₃Is H, alkyl or substituted alkyl, alkenyl or substituted alkenyl, alkynyl or substituted alkynyl, cycloalkyl or substituted cycloalkyl, cycloalkenyl or substituted cycloalkenyl, aryl or substituted aryl or heteroaryl or substituted heteroaryl; wherein R is₃Optionally substituted by one or more R₁Substitution;

R₁independently for each occurrence is H, halogen, aryl OR substituted aryl, heteroaryl OR substituted heteroaryl, CN, OR_A、SR_A、NR_AR_B、-NR_A(CH₂)_OOR_B、-N((CH₂)_OOR_A)((CH₂)_OOR_B)-C(＝O)R_A、-C(＝O)OR_A、-OC(＝O)R_A、-OC(＝O)(C₁-C₆alkyl-R_H)、-OC(＝O)(CH₂)_oOR_A、-C(＝O)NR_AR_B、-NR_AC(＝O)R_B、-N(C(＝O)R_A)(C(＝O)R_B)、-N(C(＝O)(C₁-C₆alkyl-R_H))(C(＝O)(C₁-C₆alkyl-R_H))、-N(C(＝O)(C₁-C₆alkyl-R_H))₂、-NR_AC(＝O)(CH₂)_OOR_B、-NR_AC(＝O)(CH₂)_OOR_B、-N(C(＝O)(CH₂)_OOR_B)₂、-NR_AC(＝O)(CH₂)_ONR_AR_B、-NR_A(CH₂)_OC(＝O)OR_B、-N((CH₂)_OC(＝O)OR_A)((CH₂)_OC(＝O)OR_B)、-NR_A(CH₂)_OC(＝O)NR_AR_B、-N((CH₂)_OC(＝O)NR_AR_B)((CH₂)_OC(＝O)NR_AR_B)、-NR_A(CH₂)_OC(＝O)NR_A(CH₂)_OOR_B、-C(＝O)N((CH₂)_OOR_B)₂、-N((CH₂)_OC(＝O)NR_A(CH₂)_OOR_B)((CH₂)_OC(＝O)NR_A(CH₂)_OOR_B)、-C(＝O)NR_A(CH₂)_OOR_B、-C(＝O)NR_A(CH₂)_OOR_B、-C(＝O)NR_A(CH₂)_ONR_AR_B、-N＝CR_A-NR_AR_B、-NR_B-C(＝NH)-NR_AR_B、O(CH₂)_mOR_A、O(CH₂)_mCOOR_A、O(CH₂)_mCONR_AR_B、O(CH₂)_mCONR_A(CH₂)_mOR_A、O(CH₂)_mO(CH₂)_mOR_A、O(CH₂)_mNR_AR_B、O(CH₂)_mO(CH₂)_mNR_AR_B、NR_C(CH₂)_mNR_AR_B、NR_C(CH₂)_mNR_C(CH₂)_mNR_AR_B、Wherein said aryl or heteroaryl is optionally substituted by a substituent selected from the group consisting of halogen, hydroxy, (C)₁-C₆) Alkyl group, (CH)₂)_pOR_A、(CH₂)_pNR_AR_B、(CH₂)_pC(＝O)NR_AR_BAnd (CH)₂)_pC(＝O)OR_AAnd may be the same or different and is substituted by one or more groups;

R₇is composed of

Each R₅Independently is H, alkyl or substituted alkyl, alkenyl or substituted alkenyl, alkynyl or substituted alkynyl, cycloalkyl or substituted cycloalkyl, cycloalkenyl or substituted cycloalkenyl or aryl or substituted aryl;

R_Aand R_BIndependently for each occurrence:

hydrogen;

(C₁-C₆) Alkyl optionally substituted by one or more groups R which may be the same or different_DSubstitution;

(C₂-C₆) Alkenyl or (C)₂-C₆) An alkynyl group;

optionally is (C)₁-C₆) Alkyl substituted (C)₃-C₇) A cycloalkyl group;

optionally substituted by a group selected from halogen, -O (C)₁-C₆) Alkyl, -C (═ O) O (C)₁-C₆) Phenyl or benzyl of alkyl, amino, alkylamino and dialkylamino, which may be substituted identically or differently by 1 to 5 radicals;

or may be a saturated or unsaturated heterocyclic ring containing 5 or 6 ring atoms and 1 to 3 heteroatoms, which may be the same or different, selected from nitrogen, sulphur and oxygen;

or R_AAnd R_BTogether with the nitrogen atom to which they are attached form a saturated or unsaturated heterocyclic ring containing from 3 to 7 ring atoms, which may optionally contain a substituent selected from nitrogen,Oxygen and sulfur, and may be optionally substituted with 1 to 4 groups, which may be the same or different, selected from alkyl, phenyl and benzyl;

R_Ceach occurrence is independently hydrogen or (C)₁-C₆) An alkyl group;

R_Dindependently at each occurrence is halogen, hydroxy, O (C)₁-C₄) Alkyl, C (═ O) (C)₁-C₄) Alkyl, C (═ O) O (C)₁-C₄) Alkyl orWherein Z is CH₂、O、S、NH、NCH₃NEt, N-isopropyl, N-neopentyl, N-CH₂CH₂OH or N-CH₂CH₂OMe；

R_GEach occurrence is independently R_A、OR_A、SR_A、NR_AR_B、–(CH₂)_oR_A、–(CH₂)_oC(＝O)OR_A、–(CH₂)_oC(＝O)NR_AR_B、C(＝O)OR_A、OC(＝O)R_A、NR_AC(＝O)R_B、NR_AC(＝O)(CH₂)_OOR_A、C(＝O)O(CH₂)_OOR_A、C(＝O)OR_B、C(＝O)NR_AR_B、C(＝O)NR_A(CH₂)_OOR_B、C(＝O)N((CH₂)_OOR_A)((CH₂)_OOR_B)、C(＝O)N((CH₂)_OC(＝O)OR_A)((CH₂)_OC(＝O)OR_B)、C(＝O)N((CH₂)_ONR_AR_B)((CH₂)_ONR_AR_B)、C(＝O)N((CH₂)_OOC(＝O)(CH₂)_OOR_A)((CH₂)_OOC(＝O)(CH₂)_OOR_B)、C(＝O)N((CH₂)_ONR_AC(＝O)(CH₂)_OOR_B)((CH₂)_ONR_AC(＝O)(CH₂)_OOR_B)、C(＝O)NR_A(CH₂)_ONR_AR_B、、C(＝O)NR_A(CH₂)_OOC(＝O)R_B、C(＝O)NR_A(CH₂)_OC(＝O)OR_B、C(＝O)NR_A(CH₂)_OC(＝O)NR_AR_B、C(＝O)NR_A(CH₂)_OOC(＝O)(CH₂)_OOR_BOr

R_HIndependently for each occurrence is halogen;

z' is independently CH at each occurrence₂、O、S、NR_A、N(CH₂)_oOR_A、N(CH₂)_oNR_AR_B、N(CH₂)_oCOOR_A、N(CH₂)_oOC(＝O)R_A、N(CH₂)_oCONR_AR_B、N(CH₂)_oNR_AC(＝O)R_BOr N (CH)₂)_oOC(＝O)(CH₂)_oOR_A；

o is independently at each occurrence 0,1, 2,3, 4, 5 or 6;

p is independently at each occurrence an integer of 0,1, 2,3, 4, or 5; and is

m is independently at each occurrence an integer of 1, 2,3, 4 or 5.

In some embodiments, x is 0. In other embodiments, x is 1. In some embodiments, R₈Is (C)₁-C₁₂) Alkyl radical、(C₂-C₁₂) Alkenyl, (C)₂-C₁₂) Alkynyl, (C)₃-C₁₂) Cycloalkyl or phenyl or CH₂-phenyl, optionally substituted by a group selected from halogen, hydroxy, (C)₁-C₆) Alkyl, which may be the same or different, is substituted with one or more groups. In some embodiments, R₈Is (C)₁-C₁₂) An alkyl group. In other embodiments, R₈Is (C)₁-C₆) A linear alkyl group. In other embodiments, R₈Is (C)₇-C₁₂) A linear alkyl group. In other embodiments, R₈Is (C)₄-C₆) A linear alkyl group. In other embodiments, R₈Is (C)₆-C₈) A linear alkyl group. In some embodiments, R₈Is- (CH)₂)_3-11-an alkyl chain.

In some embodiments, R₈Is (C)₂-C₁₂) An alkenyl group. In other embodiments, R₈Is (C)₂-C₆) A linear alkenyl group. In other embodiments, R₈Is (C)₇-C₁₂) A linear alkenyl group. In other embodiments, R₈Is (C)₄-C₆) A linear alkenyl group. In other embodiments, R₈Is (C)₆-C₈) A linear alkenyl group.

In some embodiments, R₈Is (C)₂-C₁₂) Alkynyl. In other embodiments, R₈Is (C)₂-C₆) Linear alkynyl groups. In other embodiments, R₈Is (C)₇-C₁₂) Linear alkynyl groups. In other embodiments, R₈Is (C)₄-C₆) Linear alkynyl groups. In other embodiments, R₈Is (C)₆-C₈) Linear alkynyl groups.

In some embodiments, R₁Is H. In other embodiments, R₁Is a halogen. In other embodiments, R₁Selected from the group consisting of: H. OR (OR)_A、SR_A、NR_AR_B、-C(＝O)R_A、-C(＝O)OR_A、-C(＝O)NR_AR_B、-NR_AC(＝O)R_B、-NR_AC(＝O)(CH₂)_OR_B、-NR_AC(＝O)(CH₂)_OOR_B、-NR_AC(＝O)(CH₂)_ONR_AR_B、-C(＝O)NR_A(CH₂)_OR_B、-C(＝O)NR_A(CH₂)_OOR_Band-C (═ O) NR_A(CH₂)_ONR_AR_B. In other embodiments, R₁Selected from the group consisting of: o (CH)₂)_mOR_A、O(CH₂)_mO(CH₂)_mOR_A、O(CH₂)_mNR_AR_B、O(CH₂)_mO(CH₂)_mNR_AR_B、NR_C(CH₂)_mNR_AR_BAnd NR_C(CH₂)_mNR_C(CH₂)_mNR_AR_B。

In certain embodiments, the compounds disclosed herein have the structure of formula (II) or (III):

or a pharmaceutically acceptable salt thereof, wherein:

x is 0 or 1;

y is H OR OR₅(ii) a Wherein R is₅Is H or methyl;

m 'and n' are each independently 0,1, 2,3, 4, 5,6, 7,8, 9, 10, 11, or 12;

R_A’and R_B’Independently for each occurrence:

hydrogen;

(C₁-C₆) Alkyl optionally substituted by one or more groups R which may be the same or different_DSubstitution;

(C₂-C₆) Alkenyl or (C)₂-C₆) An alkynyl group;

optionally is (C)₁-C₆) Alkyl substituted (C)₃-C₇) A cycloalkyl group;

optionally substituted by a group selected from halogen, -O (C)₁-C₆) Alkyl, -C (═ O) O (C)₁-C₆) Phenyl or benzyl of alkyl, amino, alkylamino and dialkylamino, which may be substituted identically or differently by 1 to 5 radicals;

or R_A’And R_B’Together with the nitrogen atom to which they are attached form a saturated or unsaturated heterocyclic ring containing from 3 to 7 ring atoms, which may optionally contain another heteroatom selected from nitrogen, oxygen and sulfur and may optionally be substituted by 1 to 4 groups, which may be the same or different, selected from alkyl, phenyl and benzyl; and is

R_DIndependently at each occurrence is halogen, hydroxy, O (C)₁-C₄) Alkyl, C (═ O) (C)₁-C₄) Alkyl, C (═ O) O (C)₁-C₄) Alkyl orWherein Z is CH₂、O、S、NH、NCH₃NEt, N-isopropyl, N-neopentyl, N-CH₂CH₂OH or N-CH₂CH₂OMe。

In certain embodiments, the compounds disclosed herein have the structure of formula (IV) or (V):

(ii) a Wherein x is 0 or 1; y is H OR OR₅(ii) a Wherein R is₅Is H or methyl; r_1’Each occurrence is independently H, halogen, aryl OR substituted aryl, heteroaryl OR substituted heteroaryl, OR_A、SR_A、NR_AR_B、-NR_A(CH₂)_OOR_B、-N((CH₂)_OOR_A)((CH₂)_OOR_B)-C(＝O)R_A、-C(＝O)OR_A、-OC(＝O)R_A、-OC(＝O)(C₁-C₆alkyl-R_H)、-OC(＝O)(CH₂)_oOR_A、-C(＝O)NR_AR_B、-NR_AC(＝O)R_B、-N(C(＝O)R_A)(C(＝O)R_B)、-N(C(＝O)(C₁-C₆alkyl-R_H))(C(＝O)(C₁-C₆alkyl-R_H))、-N(C(＝O)(C₁-C₆alkyl-R_H))₂、-NR_AC(＝O)(CH₂)_OOR_B、-NR_AC(＝O)(CH₂)_OOR_B、-N(C(＝O)(CH₂)_OOR_B)₂、-NR_AC(＝O)(CH₂)_ONR_AR_B、-NR_A(CH₂)_OC(＝O)OR_B、-N((CH₂)_OC(＝O)OR_A)((CH₂)_OC(＝O)OR_B)、-NR_A(CH₂)_OC(＝O)NR_AR_B、-N((CH₂)_OC(＝O)NR_AR_B)((CH₂)_OC(＝O)NR_AR_B)、-NR_A(CH₂)_OC(＝O)NR_A(CH₂)_OOR_B、-C(＝O)N((CH₂)_OOR_B)₂、-N((CH₂)_OC(＝O)NR_A(CH₂)_OOR_B)((CH₂)_OC(＝O)NR_A(CH₂)_OOR_B)、-C(＝O)NR_A(CH₂)_OOR_B、-C(＝O)NR_A(CH₂)_OOR_B、-C(＝O)NR_A(CH₂)_ONR_AR_B、-N＝CR_A-NR_AR_B、-NR_B-C(＝NH)-NR_AR_B、O(CH₂)_mOR_A、O(CH₂)_mCOOR_A、O(CH₂)_mCONR_AR_B、O(CH₂)_mCONR_A(CH₂)_mOR_A、O(CH₂)_mO(CH₂)_mOR_A、O(CH₂)_mNR_AR_B、O(CH₂)_mO(CH₂)_mNR_AR_B、NR_C(CH₂)_mNR_AR_B、NR_C(CH₂)_mNR_C(CH₂)_mNR_AR_B、Wherein said aryl or heteroaryl is optionally substituted by a substituent selected from the group consisting of halogen, hydroxy, (C)₁-C₆) Alkyl group, (CH)₂)_pOR_A、(CH₂)_pNR_AR_B、(CH₂)_pC(＝O)NR_AR_BAnd (CH)₂)_pC(＝O)OR_AMay be the same or different.

In certain embodiments, R_1’Is OR_A、OCOCH₂OR_A、SR_A、NHR_A、N(R_A)₂Or NHCOCH₂OR_A. In certain embodiments, R_1’Is OAc, OCOCH₂Cl、OCOCH₂CH₃、OCOCHMe₂、OCOCMe₃、OCOCH＝CH₂、NHCH₂CH₂OH、NHCH₂CH₂OMe、N(CH₂CH₂OH)₂、N(CH₂CH₂OMe)₂、NHCH₂CHMe₂、NHCH₂CMe₂、NHAc、NHCH₂COOH、NHCH₂COOCH₃、NMeCH₂COOH、NMeCH₂COOCH₃、NHCH₂CONH₂、NHCH₂CONHMe、NHCH₂CONMe₂、NHCH₂CONHCH₂CH₂OH、NHCH₂CONHCH₂CH₂OMe、NMeCH₂CONH₂、NMeCH₂CONHMe、NMeCH₂CONMe₂、N(CH₂COOH)₂、N(CH₂CONH₂)₂、N(CH₂CONHMe)₂、N(CH₂CONMe₂)₂、N(CH₂CONHCH₂CH₂OH)₂、N(CH₂CONHCH₂CH₂OMe)₂、NHCOCH₂Cl、NHCOCH₂CH₃、NHCOCHMe₂、NHCOCMe₃、NHCOCH＝CH₂、N(COCH₂Cl)₂、N(COCH₂CH₃)₂、N(COCHMe₂)₂、N(COCMe₃)₂Or N (COCH ═ CH)₂)₂。

In certain embodiments, R_A’And R_B’Each independently of the others is H, Me, Et, n-propyl, isopropyl, isobutyl, neopentyl, cyclopentyl, cyclohexyl, CH₂CH₂OH、CH₂CH₂OMe、Wherein Z is CH₂、O、S、NH、NCH₃NEt, N-isopropyl, N-neopentyl, N-CH₂CH₂OH or N-CH₂CH₂And OMe (organic chemical engineering) is adopted. In certain embodiments, R_A’And R_B’Each being Me. In certain embodiments, R_A’And R_B’Each is Et.

In certain embodiments, the compounds disclosed herein have the structure of formula (VI):

wherein

x is 0 or 1;

w is CH₂O or S;

y is H OR OR₅(ii) a Wherein R is₅Is H or methyl;

m 'and n' are each independently 0,1, 2,3, 4, 5,6, 7,8, 9, or 10; and is

R₁Each occurrence is independently H, halogen, aryl OR substituted aryl, heteroaryl OR substituted heteroaryl, OR_A、SR_A、NR_AR_B、-C(＝O)R_A、-C(＝O)OR_A、-C(＝O)NR_AR_B、-NR_AC(＝O)R_B、-NR_AC(＝O)(CH₂)_OR_B、-NR_AC(＝O)(CH₂)_OOR_B、-NR_AC(＝O)(CH₂)_ONR_AR_Bor-C (═ O) NR_A(CH₂)_OR_B。

In certain embodiments, R₁Selected from the group consisting of: H. OR (OR)_A、SR_A、NR_AR_B、-C(＝O)R_A、-C(＝O)OR_A、-C(＝O)NR_AR_B、-NR_AC(＝O)R_B、-NR_AC(＝O)(CH₂)_OR_B、-NR_AC(＝O)(CH₂)_OOR_B、-NR_AC(＝O)(CH₂)_ONR_AR_B、-C(＝O)NR_A(CH₂)_OR_B、-C(＝O)NR_A(CH₂)_OOR_Band-C (═ O) NR_A(CH₂)_ONR_AR_B。

In certain embodiments, R₁Selected from the group consisting of: o (CH)₂)_mOR_A、O(CH₂)_mO(CH₂)_mOR_A、O(CH₂)_mNR_AR_B、O(CH₂)_mO(CH₂)_mNR_AR_B、NR_C(CH₂)_mNR_AR_BAnd NR_C(CH₂)_mNR_C(CH₂)_mNR_AR_B。

In certain embodiments, R₁Selected from the group consisting of: OR (OR)_A、SR_A、NR_AR_B、-C(＝O)R_A、-C(＝O)OR_Aand-C (═ O) NR_AR_B. In some embodiments, R₁Is OH, OMe, OEt, O-isopropyl, O-isobutyl, O-neopentyl, O-cyclopentyl, O-cyclohexyl, SH, SMe, S-isopropyl, S-isobutyl, S-neopentyl, S-cyclopentyl, S-cyclohexyl,In certain embodiments, R₁Is composed ofWherein Z isCH₂、O、S、NH、NCH₃NEt, N-isopropyl, N-neopentyl, N-CH₂CH₂OH or N-CH₂CH₂And OMe (organic chemical engineering) is adopted. In some embodiments, described herein are R_AAnd R_B. In other embodiments, R_AAnd R_BEach is H, Me, Et, isopropyl, isobutyl, cyclopentyl, or cyclohexyl. In certain embodiments, R₁Is a hydroxyl group. In certain other embodiments, R₁Is C (═ O) OR_A。

In other embodiments, R₁Selected from the group consisting of: H. OH, OMe, OEt, O-isopropyl, O-isobutyl, O-neopentyl, O-cyclopentyl, O-cyclohexyl, OCH₂CH₂OH、OCH₂CH₂OCH₃、OCH₂COOH、OCH₂COOCH₃、OCH₂CONH₂、OCH₂CONHMe、OCH₂CONMe₂、OCH₂CONHCH₂CH₂OH、OCH₂CONHCH₂CH₂OMe、OAc、OOCCH₂Cl、OOCCH₂OR_A、OOCCH₂CH₃、OOCCHMe₂、OOCCMe₃And OC (O) CCH (CH)₂。

In other embodiments, R₁Selected from the group consisting of:

in other embodiments, R₁Is SH, SMe, SEt, S-isopropyl, S-isobutyl, S-neopentyl, O-cyclopentyl or S-cyclohexyl.

In other embodiments, R₁Selected from the group consisting of: NH (NH)₂、NHCH₃、NHCH₂CH₃、NHCH₂CHOH、NHCH₂CH₂OMe、NMe₂、NEt₂、NHCH₂CHMe₂、NHCH₂CMe₃、NHAc、NHCH₂COOH、NHCH₂COOCH₃、NMeCH₂COOH、NMeCH₂COOCH₃、NHCH₂CONH₂、NHCH₂CONHMe、NHCH₂CONMe₂、NHCH₂CONHCH₂CH₂OH、NHCH₂CONHCH₂CH₂OMe、NMeCH₂CONH₂、NMeCH₂CONHMe、NMeCH₂CONMe₂、N(CH₂COOH)₂、N(CH₂CONH₂)₂、N(CH₂CONHMe)₂、N(CH₂CONMe₂)₂、N(CH₂CONHCH₂CH₂OH)₂、N(CH₂CONHCH₂CH₂OMe)₂、NHCOCH₂Cl、NHCOCH₂OR_A、NHCOCH₂CH₃、NHCOCHMe₂、NHCOCMe₃、NHCOCH＝CH₂、N(COCH₂Cl)₂、N(COCH₂OR_A)₂、N(COCH₂CH₃)₂、N(COCHMe₂)₂、N(COCMe₃)₂And N (COCH ═ CH)₂)₂。

In other embodiments, R₁Is composed ofWherein Z is CH₂O, S, NH, NMe, NEt, N-isopropyl, N-neopentyl, N-CH₂CH₂OH、NCH₂CH₂OCH₃、NCH₂CH₂OCH₃、NCH₂COOH、NCH₂COOMe、N-CH₂CONH₂、NCH₂CONHMe or NCH₂CONMe₂。

In other embodiments, R₁Is selected from the group consisting ofThe group consisting of:

in other embodiments, R₁Selected from the group consisting of:

wherein Z' is independently at each occurrence CH₂、O、S、NR_A、N(CH₂)_oOR_A、N(CH₂)_oNR_AR_B、N(CH₂)_oCOOR_A、N(CH₂)_oOC(＝O)R_A、N(CH₂)_oCONR_AR_B、N(CH₂)_oNR_AC(＝O)R_BOr N (CH)₂)_oOC(＝O)(CH₂)_oOR_A。

In other embodiments, R₁is-COOH, -COOMe, -COOEt, -CONH₂、-CONHMe、-CONMe₂、-CONHEt、-CONEt₂、-CONHCH₂CH₂OH、-CONHCH₂CH₂OMe、-CON(CH₂CH₂OH)₂、-CON(CH₂CH₂OMe)₂or-CONMe₂。

In other embodiments, R₁Selected from the group consisting of:

(ii) a Wherein Z is CH₂、O、S、NH、NCH₃NEt, N-isopropyl, N-neopentyl, N-CH₂CH₂OH or N-CH₂CH₂OMe。

In other embodiments, R₁Is composed ofIn some embodiments, R_GIs OH, OMe, OAc, NH₂、NHMe、NHAc、NMe₂、NEt₂、NHCH₂CMe₃。

In other embodiments, R₁Selected from the group consisting of:

in other embodiments, R₁Is OR_A、OCOCH₂OR_A、SR_A、NHR_A、N(R_A)₂Or NHCOCH₂OR_A。

In other embodiments, R₁Is OAc, OCOCH₂Cl、OCOCH₂CH₃、OCOCHMe₂、OCOCMe₃、OCOCH＝CH₂、NHCH₂CH₂OH、NHCH₂CH₂OMe、N(CH₂CH₂OH)₂、N(CH₂CH₂OMe)₂、NHCH₂CHMe₂、NHCH₂CMe₂、NHAc、NHCH₂COOH、NHCH₂COOCH₃、NMeCH₂COOH、NMeCH₂COOCH₃、NHCH₂CONH₂、NHCH₂CONHMe、NHCH₂CONMe₂、NHCH₂CONHCH₂CH₂OH、NHCH₂CONHCH₂CH₂OMe、NMeCH₂CONH₂、NMeCH₂CONHMe、NMeCH₂CONMe₂、N(CH₂COOH)₂、N(CH₂CONH₂)₂、N(CH₂CONHMe)₂、N(CH₂CONMe₂)₂、N(CH₂CONHCH₂CH₂OH)₂、N(CH₂CONHCH₂CH₂OMe)₂、NHCOCH₂Cl、NHCOCH₂CH₃、NHCOCHMe₂、NHCOCMe₃、NHCOCH＝CH₂、N(COCH₂Cl)₂、N(COCH₂CH₃)₂、N(COCHMe₂)₂、N(COCMe₃)₂Or N (COCH ═ CH)₂)₂。

In other embodiments, R₁Is aryl or heteroaryl, said aryl or heteroaryl being optionally substituted by a group selected from halogen, hydroxy, (C)₁-C₆) Alkyl, (C)₃-C₇) Cycloalkyl, SR_A、(CH₂)_pOR_A、(CH₂)_pNR_AR_B、(CH₂)_pC(＝O)R_A、(CH₂)_pC(＝O)NR_AR_BAnd (CH)₂)_pC(＝O)OR_AAnd may be the same or different and is substituted by one or more groups; wherein p is 0,1, 2,3, 4, 5, 6; and herein describes R_AAnd R_BEach occurrence of (a). In some embodiments, R₁Is thatWherein Rx is H, (C)₁-C₆) Alkyl or (C)₃-C₇) A cycloalkyl group; ry is H, (C)₁-C₆) Alkyl, (C)₃-C₇) Cycloalkyl, OR_A、SR_A、NR_AR_B、-C(＝O)R_A、-C(＝O)OR_Aor-C (═ O) NR_AR_B(ii) a And t is 1, 2,3 or 4. In some embodiments, Rx is H or Me; and Ry is-C (═ O) OR_Aor-C (═ O) NR_AR_B. In some embodiments, R₁Is composed ofWherein Z is CH₂、O、S、NH、NCH₃NEt, N-isopropyl, N-neopentyl, N-CH₂CH₂OH or N-CH₂CH₂OMe。

In other embodiments, R_AAnd R_BEach independently of the others is H, Me, Et, isopropyl, isobutyl, neopentyl, cyclopentyl, cyclohexyl, CH₂CH₂OH、CH₂CH₂OMe orWherein Z is CH₂、O、S、NH、NCH₃NEt, N-isopropyl, N-neopentyl, N-CH₂CH₂OH or N-CH₂CH₂And OMe (organic chemical engineering) is adopted. In some embodiments, R_AAnd R_BEach independently is H, Me, Et, isopropyl, isobutyl, cyclopentyl, or cyclohexyl. In some embodiments, R_AAnd R_BEach occurrence is independently H or (C)₁-C₆) An alkyl group.

In some embodiments, R₃Is H, (C)₁-C₁₂) Alkyl, (C)₂-C₁₂) Alkenyl, (C)₂-C₁₂) Alkynyl, (C)₃-C₁₂) Cycloalkyl or phenyl or CH₂-phenyl, optionally substituted by a group selected from halogen, hydroxy, (C)₁-C₆) Alkyl, which may be the same or different, is substituted with one or more groups. In some embodiments, R₃Is (C)₁-C₁₂) An alkyl group. In other embodiments, R₃Is (C)₁-C₆) A linear alkyl group. In other embodiments, R₃Is (C)₇-C₁₂) A linear alkyl group. In other embodiments, R₃Is (C)₄-C₆) A linear alkyl group. In other embodiments, R₃Is (C)₆-C₈) A linear alkyl group. In certain embodiments, R₃Is (C)₇-C₁₀) An alkyl group. In certain other embodiments, R₃Is (C)₇-C₈) An alkyl group. In other embodiments, R₃Is (C)₇-C₁₀) A linear alkyl group. In other embodiments, R₃Is (C)₇-C₈) A linear alkyl group. In some embodiments, R₃Is- (CH)₂)_3-11-an alkyl chain.

In some embodiments, R₃Is (C)₂-C₁₂) An alkenyl group. In other embodiments, R₃Is (C)₂-C₆) A linear alkenyl group. In other embodiments, R₃Is (C)₇-C₁₂) A linear alkenyl group. In other embodiments, R₃Is (C)₄-C₆) A linear alkenyl group. In other embodiments, R₃Is (C)₆-C₈) A linear alkenyl group.

In some embodiments, R₃Is (C)₂-C₁₂) Alkynyl. In other embodiments, R₃Is (C)₂-C₆) Linear alkynyl groups. In other embodiments, R₃Is (C)₇-C₁₂) Linear alkynyl groups. In other embodiments, R₃Is (C)₄-C₆) Linear alkynyl groups. In other embodiments, R₃Is (C)₆-C₈) Linear alkynyl groups. In any of the embodiments described herein, R₃It may be 4-hydroxybutyl.

In certain embodiments, R₂Is ethyl. In certain embodiments, R₇Is composed ofIn certain embodiments, W is O or S. In certain embodiments described herein, W is S; x is 1; and R is₃Is 4-hydroxybutyl.

In certain embodiments, the compounds of formula I have the structures of formulae (II ') to (VI'):

or a pharmaceutically acceptable salt thereof, wherein:

each x is 0 or 1; each W is independently O, S or CH₂，

R₁Each occurrence is independently H, halogen, aryl, heteroaryl, OR_A、SR_A、NR_AR_B、-C(＝O)R_A、-C(＝O)OR_A、-C(＝O)NR_AR_B、-NR_AC(＝O)R_B、-NR_AC(＝O)(CH₂)_OR_B、-NR_AC(＝O)(CH₂)_OOR_B、-NR_AC(＝O)(CH₂)_ONR_AR_B、-C(＝O)NR_A(CH₂)_OR_B、-N＝CR_A-NR_AR_B、-NR_B-C(＝NH)-NR_AR_B、-C(＝O)NR_A(CH₂)_OOR_B、-C(＝O)NR_A(CH₂)_ONR_AR_B、O(CH₂)_mOR_A、O(CH₂)_mO(CH₂)_mOR_A、O(CH₂)_mNR_AR_B、O(CH₂)_mO(CH₂)_mNR_AR_B、NR_C(CH₂)_mNR_AR_BOr NR_C(CH₂)_mNR_C(CH₂)_mNR_AR_BWherein said aryl or heteroaryl is optionally substituted by a substituent selected from the group consisting of halogen, hydroxy, (C)₁-C₆) Alkyl group, (CH)₂)_pOR_A、(CH₂)_pNR_AR_B、(CH₂)_pC(＝O)NR_AR_BAnd (CH)₂)_pC(＝O)OR_AAnd may be the same or different and is substituted by one or more groups;

each R₃Independently is (C)₁-C₆) Alkyl, (C)₇-C₁₂) Alkyl, (C)₂-C₆) Alkenyl, (C)₇-C₁₂) Alkenyl, (C)₂-C₆) Alkynyl or (C)₇-C₁₂) Alkynyl, aryl or heteroaryl, all of which may optionally be substituted by one or more radicals R which may be the same or different₁Substitution;

each R₅Independently are:

H；

(C₁-C₆) Alkyl optionally substituted by one or more groups R which may be the same or different₆Substitution;

(C₂-C₆) Alkenyl, which are optionally substituted by radicals which may be identical or different and are each selected from halogen, hydroxy, (C)₁-C₆) Alkyl, aryl (e.g. phenyl), (CH)₂)_pOR_A、(CH₂)_mOH、(CH₂)_mO(CH₂)_mOH、(CH₂)_mNR_AR_B、(CH₂)_mO(CH₂)_mNR_AR_B、(CH₂)_pNR_AR_B、(CH₂)_pNR_C(CH₂)_mNR_AR_B、(CH₂)_pNR_c(CH₂)_mNR_c(CH₂)_mNR_AR_B、(CH₂)_pC(＝O)NR_AR_B、(CH₂)_pC(＝O)OR_ASubstituted with one or more groups of (a);

(C₂-C₆) Alkynyl optionally substituted with 1 or more groups which may be the same or different and are each selected from halo, hydroxy, amino, monoalkylamino and dialkylamino;

(C₃-C₇) Cycloalkyl optionally substituted with 1 or more groups which may be the same or different and are each selected from halogen, hydroxy, amino, monoalkylamino and dialkylamino;

phenyl or CH₂-phenyl, optionally substituted by radicals which may be identical or different and are each chosen from halogen, hydroxy, (C)₁-C₆) Alkyl group, (CH)₂)_pOR_A、(CH₂)_pNR_AR_B、(CH₂)_pC(＝O)NR_AR_B、(CH₂)_pC(＝O)OR_ASubstituted with one or more groups of (a);

R₆independently at each occurrence is halogen, hydroxy, aryl (e.g. phenyl), S (C)₁-C₆) Alkyl, SR_A、OR_A、O(CH₂)_mOR_A、O(CH₂)_mO(CH₂)_mOR_A、C(＝O)OR_A、C(＝O)NR_AR_B、NR_AR_B、O(CH₂)_mNR_AR_B、O(CH₂)_mO(CH₂)_mNR_AR_B、NR_C(CH₂)_mNR_AR_BOr NR_C(CH₂)_mNR_C(CH₂)_mNR_AR_BWherein said aryl or phenyl is optionally substituted withIdentical or different and are each selected from halogen, hydroxy, (C)₁-C₆) Alkyl group, (CH)₂)_pOR_A、(CH₂)_pNR_AR_B、(CH₂)_pC(＝O)NR_AR_BAnd (CH)₂)_pC(＝O)OR_ASubstituted with one or more groups of (a);

R_Aand R_BIndependently for each occurrence:

hydrogen;

(C₁-C₆) Alkyl optionally substituted by one or more groups R which may be the same or different_DSubstitution;

(C₂-C₆) Alkenyl or (C)₂-C₆) An alkynyl group;

optionally is (C)₁-C₆) Alkyl substituted (C)₃-C₇) A cycloalkyl group;

optionally substituted by one or more substituents which may be the same or different and are each selected from halogen, -O (C)₁-C₆) Alkyl, -C (═ O) O (C)₁-C₆) Phenyl substituted with 1 to 5 groups of alkyl, amino, alkylamino and dialkylamino;

or may be a saturated or unsaturated heterocyclic ring containing 5 or 6 ring atoms and 1 to 3 heteroatoms which may be the same or different and are each selected from nitrogen, sulphur and oxygen;

or R_AAnd R_BTogether with the nitrogen atom to which they are attached form a saturated or unsaturated heterocyclic ring containing from 3 to 7 ring atoms, which may optionally contain another heteroatom each selected from nitrogen, oxygen and sulfur and may optionally be substituted with from 1 to 4 groups which may be the same or different and each selected from alkyl, phenyl and benzyl;

or R_AAnd R_BTogether with the nitrogen atom to which they are attached form-N ═ CH-NR_FR_F’、-N＝CMe-NR_FR_F’or-NR_FC(＝NH)NR_FR_F’；

R_CEach occurrence is independently hydrogen or (C)₁-C₆) An alkyl group;

R_Dindependently at each occurrence is halogen, hydroxy, O (C)₁-C₄) Alkyl, C (═ O) (C)₁-C₄) Alkyl, C (═ O) O (C)₁-C₄) An alkyl group;

R_Fand R_F’Independently at each occurrence, is hydrogen, (C)₁-C₆) Alkyl, phenyl, benzyl, or R_FAnd R_F’Together with the nitrogen atom to which they are attached form a saturated or unsaturated heterocyclic ring containing from 3 to 7 ring atoms, which may optionally contain another heteroatom each selected from nitrogen, oxygen and sulfur and may optionally be substituted with from 1 to 4 groups which may be the same or different and each selected from alkyl, phenyl and benzyl;

p is an integer 0,1, 2,3, 4, 5 or 6;

m' is 0,1, 2,3, 4, 5,6, 7,8 or 9;

m is an integer 1, 2,3, 4, 5 or 6; and is

n is an integer 1, 2,3, 4, 5 or 6.

In any of the embodiments described herein, x is 0 or 1. In any of the embodiments described herein, x can be 0. In any of the embodiments described herein, x can be 1. In certain embodiments, W is O. In certain other embodiments, W is S. In other embodiments, W is CH₂。

In certain embodiments, m' is 1. In certain other embodiments, m' is 2. In other embodiments, m' is 3. In other embodiments, m' is 4 or 5. In other embodiments, m' is 6,7, 8, or 9.

In certain embodiments, m is 1. In certain other embodiments, m is 2. In other embodiments, m is 3. In other embodiments, m is 4 or 5.

In certain embodiments, p is 0. In certain other embodiments, p is 1. In other embodiments, m is 2. In other embodiments, m is 3,4, or 5.

In certain embodiments, R₃Is composed ofIn certain embodiments, R₃Is H, methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, tert-butyl, CH₂CMe₃Phenyl, CH₂-phenyl group,

In certain embodiments, R₃Is- (CH)₂)_nNR_AR_BWherein n is an integer 2,3, 4, 5 or 6, or an integer 7,8, 9, 10, 11 or 12; and wherein R_AAnd R_BIndependently for each occurrence hydrogen; (C)₁-C₄) Alkyl optionally substituted by one or more groups R which may be the same or different_DIs substituted in which R_DIndependently at each occurrence is halogen, hydroxy, O (C)₁-C₄) Alkyl, C (═ O) (C)₁-C₄) Alkyl, C (═ O) O (C)₁-C₄) An alkyl group; or R_AAnd R_BTogether with the nitrogen atom to which they are attached form a saturated or unsaturated heterocyclic ring containing from 3 to 7 ring atoms, which may optionally contain another heteroatom selected from nitrogen, oxygen and sulfur and may optionally be selected from (C)₁-C₄) Of alkyl, phenyl and benzyl groups,May be substituted by 1 to 4 groups which may be the same or different.

In certain embodiments, R₃Is- (CH)₂)_nNR_AR_BWherein n is an integer 2,3, 4, 5 or 6, or an integer 7,8, 9, 10, 11 or 12; and wherein R_AAnd R_BTogether with the nitrogen atom to which they are attached form a saturated or unsaturated heterocyclic ring containing from 3 to 7 ring atoms, which may optionally contain another heteroatom selected from nitrogen, oxygen and sulfur and may optionally be selected from (C)₁-C₄) Alkyl, phenyl and benzyl, which may be substituted by 1 to 4 groups, which may be the same or different.

In certain embodiments, n is 2. In certain other embodiments, n is 3. In other embodiments, n is 4, 5, or 6. In other embodiments, n is 7 or 8. In other embodiments, n is 9 or 10. In other embodiments, n is 11 or 12.

In certain embodiments, R₃Is- (CH)₂)_nNR_AR_BWherein n is an integer of 7,8, 9, 10, 11 or 12; and wherein R_AAnd R_BIndependently for each occurrence hydrogen; (C)₁-C₄) Alkyl optionally substituted by one or more groups R which may be the same or different_DIs substituted in which R_DIndependently at each occurrence is halogen, hydroxy, O (C)₁-C₄) Alkyl, C (═ O) (C)₁-C₄) Alkyl, C (═ O) O (C)₁-C₄) An alkyl group; or R_AAnd R_BTogether with the nitrogen atom to which they are attached form a saturated or unsaturated heterocyclic ring containing from 3 to 7 ring atoms, which may optionally contain another heteroatom selected from nitrogen, oxygen and sulfur and may optionally be selected from (C)₁-C₄) The alkyl, phenyl and benzyl groups may be substituted by 1 to 4 groups which may be the same or different.

In certain embodiments, n is 7. In certain other embodiments, n is 8. In other embodiments, n is 9, 10, 11, or 12.

In certain embodiments, R₃Is 2-aminoethyl, 2-aminopropyl, 3-aminopropyl, 2-monoalkylaminoethyl, 2-monoalkylaminopropyl, 3-monoalkylaminopropyl, 2-dialkylaminoethyl, 2-dialkylaminopropyl or 3-dialkylaminopropyl, wherein the alkyl is (C)₁-C₄) An alkyl group.

In certain embodiments, R₃Is 2-aminoethyl, 2-aminopropyl, 3-aminopropyl, 2-monoalkylaminoethyl, 2-monoalkylaminopropyl, 3-monoalkylaminopropyl, 2-dialkylaminoethyl, 2-dialkylaminopropyl or 3-dialkylaminopropyl, wherein the alkyl is (C)₁-C₄) Alkyl radical, wherein R₃Is dimethylaminoethyl, diethylaminoethyl, methylethylaminoethyl, methyl-isobutylaminoethyl, ethyl-isobutylaminoethyl, methyl-tert-butylaminoethyl or ethyl-tert-butylaminoethyl.

In certain embodiments, R₃Is composed ofWherein n is an integer of 2,3, 4, 5 or 6, and m is an integer of 2,3 or 4. In certain embodiments, n is 2. In certain other embodiments, n is 3. In other embodiments, n is 4 or 5 or 6. In certain embodiments, m is 2. In certain other embodiments, m is 3. In certain other embodiments, m is 4. In certain embodiments, n is 7. In certain other embodiments, n is 8. In other embodiments, n is 9, 10, 11, or 12.

In certain embodiments, R₅Is H, (C)₁-C₆) Alkyl, (C)₂-C₆) Alkenyl, phenyl, benzyl、CH₂-S-(C₁-C₆) Alkyl radical, CH₂-O-(C₁-C₆) Alkyl, (C)₂-C₆)OR_A、(C₁-C₆) Monoalkylamino, (C)₁-C₆) A dialkylamino group or (C)₁-C₆) -cyclic amines wherein the phenyl or benzyl group is optionally selected from (C)₁-C₄) Alkyl, (C)₁-C₄) Alkoxy and halogen with 1-3 substituents; and R is_AIs H, (C)₁-C₆) Alkyl, phenyl, CH₂-phenyl, (C)₁-C₆) Alkyl OH, (CH)₂)_pO(CH₂)_mOH、(CH₂)_pO(CH₂)_mO(CH₂)_mOH、(C₁-C₆) Alkyl O (C)₁-C₄) Alkyl group, (CH)₂)_pO(CH₂)_mO(C₁-C₄) Alkyl or (CH)₂)_pO(CH₂)_mO(CH₂)_mO(C₁-C₄) An alkyl group; p is an integer 0,1, 2,3, 4 or 5; and m is an integer of 1, 2,3, 4 or 5.

In certain embodiments, R₅Is H. In certain other embodiments, R₅Is methyl. In other embodiments, R₅Is CH₂-S-(C₁-C₆) Alkyl radicals, e.g. CH₂-S-CH₃. In other embodiments, R₅Is CH₂-O-(C₁-C₆) Alkyl radicals, e.g. CH₂-O-CH₂-CH₃. In other embodiments, R₅Is (C)₂-C₆) Alkenyl radicals, e.g. CH₂-CH＝CH₂. In other embodiments, R₅Is benzyl. In other embodiments, R₅Is (C)₂-C₆) And (5) OH. In other embodiments, R₅Is (C)₁-C₆) Monoalkylamines, e.g. CH₂-NH-Me. In other embodiments, R₅Is (C)₁-C₆) Dialkylamines, e.g. CH₂-CH₂-N(Et)₂. In other embodiments, R₅Is (C)₁-C₆) Cyclic amines, e.g. CH₂-CH₂-morpholine.

In certain embodiments, R_AAnd R_BIndependently at each occurrence, H, (C)₁-C₆) Alkyl, phenyl, CH₂-phenyl, (C)₁-C₆) Alkyl OH, (CH)₂)_pO(CH₂)_mOH or (CH)₂)_pO(CH₂)_mO(CH₂)_mOH、(C₁-C₆) Alkyl O (C)₁-C₄) Alkyl group, (CH)₂)_pO(CH₂)_mO(C₁-C₄) Alkyl or (CH)₂)_pO(CH₂)_mO(CH₂)_mO(C₁-C₄) An alkyl group. In certain other embodiments, R_AAnd R_BTogether with the nitrogen atom to which they are attached form a radical selected fromWherein R is_CIs H, Me, Et, n-Pr, i-Pr, n-Bu, i-Bu, t-Bu, CH₂CMe₃、Ph、CH₂Ph、CH₂CH₂OH or CH₂CH₂O(C₁-C₄) An alkyl group.

In another aspect, the invention provides compounds of formulae (IIa) to (VIa):

or a pharmaceutically acceptable salt thereof, wherein:

each x is 0 or 1; each W is independently O, S or CH₂，

R₁Each occurrence is independently H, halogen, aryl, heteroaryl, OR_A、SR_A、NR_AR_B、-C(＝O)R_A、-C(＝O)OR_A、-C(＝O)NR_AR_B、-NR_AC(＝O)R_B、-NR_AC(＝O)(CH₂)_OR_B、-NR_AC(＝O)(CH₂)_OOR_B、-NR_AC(＝O)(CH₂)_ONR_AR_B、-C(＝O)NR_A(CH₂)_OR_B、-C(＝O)NR_A(CH₂)_OOR_B、-C(＝O)NR_A(CH₂)_ONR_AR_B、O(CH₂)_mOR_A、O(CH₂)_mO(CH₂)_mOR_A、O(CH₂)_mNR_AR_B、O(CH₂)_mO(CH₂)_mNR_AR_B、NR_C(CH₂)_mNR_AR_BOr NR_C(CH₂)_mNR_C(CH₂)_mNR_AR_BWherein said aryl or heteroaryl groups are optionally substituted by groups which may be the same or different and are each selected from halogen, hydroxy, (C)₁-C₆) Alkyl group, (CH)₂)_pOR_A、(CH₂)_pNR_AR_B、(CH₂)_pC(＝O)NR_AR_BAnd (CH)₂)_pC(＝O)OR_ASubstituted with one or more groups of (a);

each R₅Independently are:

H；

(C₁-C₆) Alkyl optionally substituted by one or more groups R which may be the same or different₆Substitution;

(C₂-C₆) Alkenyl, which are optionally substituted by radicals which may be identical or different and are each selected from halogen, hydroxy, (C)₁-C₆) Alkyl, aryl (e.g. phenyl), (CH)₂)_pOR_A、O(CH₂)_mOH、O(CH₂)_mO(CH₂)_mOH、O(CH₂)_mNR_AR_B、O(CH₂)_mO(CH₂)_mNR_AR_B、(CH₂)_pNR_AR_B、(CH₂)_pNR_C(CH₂)_mNR_AR_B、(CH₂)_pNR_C(CH₂)_mNR_c(CH₂)_mNR_AR_B、(CH₂)_pC(＝O)NR_AR_B、(CH₂)_pC(＝O)OR_ASubstituted with one or more groups of (a);

(C₂-C₆) Alkynyl optionally substituted with 1 or more groups which may be the same or different and are each selected from halo, hydroxy, amino, monoalkylamino and dialkylamino;

(C₃-C₇) Cycloalkyl optionally substituted with 1 or more groups which may be the same or different and are each selected from halogen, hydroxy, amino, monoalkylamino and dialkylamino;

phenyl or CH₂-phenyl, optionally substituted by radicals which may be identical or different and are each chosen from halogen, hydroxy, (C)₁-C₆) Alkyl group, (CH)₂)_pOR_A、(CH₂)_pNR_AR_B、(CH₂)_pC(＝O)NR_AR_B、(CH₂)_pC(＝O)OR_ASubstituted with one or more groups of (a);

R₆independently at each occurrence is halogen, hydroxy, aryl (e.g. phenyl), S (C)₁-C₆) Alkyl, SR_A、OR_A、O(CH₂)_mOR_A、O(CH₂)_mO(CH₂)_mOR_A、C(＝O)OR_A、C(＝O)NR_AR_B、NR_AR_B、O(CH₂)_mNR_AR_B、O(CH₂)_mO(CH₂)_mNR_AR_B、NR_C(CH₂)_mNR_AR_BOr NR_C(CH₂)_mNR_C(CH₂)_mNR_AR_BWherein said aryl or phenyl groups are optionally substituted by groups which may be the same or different and are each selected from halogen, hydroxy, (C)₁-C₆) Alkyl group, (CH)₂)_pOR_A、(CH₂)_pNR_AR_B、(CH₂)_pC(＝O)NR_AR_BAnd (CH)₂)_pC(＝O)OR_ASubstituted with one or more groups of (a);

R_Aand R_BIndependently for each occurrence:

hydrogen;

(C₁-C₆) Alkyl optionally substituted by one or more groups R which may be the same or different_DSubstitution;

(C₂-C₆) Alkenyl or (C)₂-C₆) An alkynyl group;

optionally is (C)₁-C₆) Alkyl substituted (C)₃-C₇) A cycloalkyl group;

optionally substituted by one or more substituents which may be the same or different and are each selected from halogen, -O (C)₁-C₆) Alkyl, -C (═ O) O (C)₁-C₆) Phenyl substituted with 1 to 5 groups of alkyl, amino, alkylamino and dialkylamino;

or may be a saturated or unsaturated heterocyclic ring containing 5 or 6 ring atoms and 1 to 3 heteroatoms which may be the same or different and are each selected from nitrogen, sulphur and oxygen;

or R_AAnd R_BTogether with the nitrogen atom to which they are attached form a saturated or unsaturated heterocyclic ring containing from 3 to 7 ring atoms, which may optionally contain another ring atom each selected from nitrogen, oxygen and sulfurA heteroatom and may be optionally substituted by 1 to 4 groups which may be the same or different and are each selected from alkyl, phenyl and benzyl;

R_Ceach occurrence is independently hydrogen or (C)₁-C₆) An alkyl group;

R_Dindependently at each occurrence is halogen, hydroxy, O (C)₁-C₄) Alkyl, C (═ O) (C)₁-C₄) Alkyl, C (═ O) O (C)₁-C₄) An alkyl group;

each p is independently an integer of 0,1, 2,3, 4, or 5;

m' is 0,1, 2,3, 4, 5,6, 7,8 or 9;

each of m and n is independently an integer of 1, 2,3, 4, or 5;

q is an integer 1, 2,3, 4, 5,6, 7,8, 9, 10 or 11.

In certain embodiments, q is 1. In certain other embodiments, q is 2. In certain other embodiments, n is independently an integer of 6,7, 8, 9, 10, or 11.

In certain embodiments, W is S. In certain other embodiments, W is O.

In certain embodiments, R₁Is hydrogen. In certain other embodiments, R₁Is (C)₁-C₆) An alkyl group. In certain embodiments, R₃Is (C)₁-C₆) An alkyl group. In certain other embodiments, R₃Is NR_CCH₂(CH₂)_pNR_AR_B。

In certain embodiments, R₅Is H, (C)₁-C₆) Alkyl, (C)₂-C₆) Alkenyl, phenyl, benzyl, CH₂-S-(C₁-C₆) Alkyl radical, CH₂-O-(C₁-C₆) Alkyl, (C)₂-C₆)OR_A、(C₁-C₆) Mono-alkylamines, (C)₁-C₆) A dialkylamine or (C)₁-C₆) -cyclic amines wherein the phenyl or benzyl group is optionally selected from (C)₁-C₄) Alkyl, (C)₁-C₄) Alkoxy and halogen with 1-3 substituents; and R is_AIs H, (C)₁-C₆) Alkyl, phenyl, CH₂-phenyl, (C)₁-C₆) Alkyl OH, (CH)₂)_pO(CH₂)_mOH、(CH₂)_pO(CH₂)_mO(CH₂)_mOH、(C₁-C₆) Alkyl O (C)₁-C₄) Alkyl group, (CH)₂)_pO(CH₂)_mO(C₁-C₄) Alkyl or (CH)₂)_pO(CH₂)_mO(CH₂)_mO(C₁-C₄) An alkyl group; p is an integer 0,1, 2,3, 4 or 5; and m is an integer of 1, 2,3, 4 or 5.

In certain other embodiments, R₅Is H, (C)₁-C₄) Alkyl, (C)₂-C₄) Alkenyl, phenyl, benzyl, CH₂-S-(C₁-C₄) Alkyl radical, CH₂-O-(C₁-C₄) Alkyl group, (CH)₂)₂OH or (CH)₂)₂O(C₁-C₄) An alkyl group. In certain embodiments, R₅Is H. In certain other embodiments, R₅Is methyl.

In certain embodiments, R_AAnd R_BIndependently at each occurrence, H, (C)₁-C₆) Alkyl, phenyl, CH₂-phenyl, (C)₁-C₆) Alkyl OH, (CH)₂)_pO(CH₂)_mOH or (CH)₂)_pO(CH₂)_mO(CH₂)_mOH、(C₁-C₆) Alkyl O (C)₁-C₄) Alkyl group, (CH)₂)_pO(CH₂)_mO(C₁-C₄) Alkyl or (CH)₂)_pO(CH₂)_mO(CH₂)_mO(C₁-C₄) An alkyl group. In certain other embodiments, R_AAnd R_BEach occurrence is independently H or (C)₁-C₆) An alkyl group. In other embodiments, R_AAnd R_BTogether with the nitrogen atom to which they are attached form a radical selected fromWherein R is_CIs H, Me, Et, n-Pr, i-Pr, n-Bu, i-Bu, t-Bu, CH₂CMe₃、Ph、CH₂Ph or CH₂CH₂OH and CH₂CH₂OR_d。

In certain embodiments, the compound is selected from the group consisting of:

and, wherein x is 0 or 1; m is 2,3, 4, 5,6, 7 or 8; y is H, OH or OMe; w is O or S; and each R_aIndependently selected from the group consisting of the moieties listed in table 1:

TABLE 1

。

In another aspect, compounds having the formula, or a pharmaceutically acceptable salt thereof,

wherein x is 1; m is 2,3, 4, 5,6, 7 or 8; y is H, OH or OMe; w is O or S; and each R_aIs that

In another aspect, compounds having the formula, or a pharmaceutically acceptable salt thereof,

wherein x is 1; m is 2,3, 4, 5,6, 7 or 8; y is H, OH or OMe; w is O or S; and each R_aIs that

In another aspect, compounds having the formula, or a pharmaceutically acceptable salt thereof,

wherein x is 1; m is 2,3, 4, 5,6, 7 or 8; y is H, OH or OMe; w is O or S; and each R_aIs that

In another aspect, compounds having the formula, or a pharmaceutically acceptable salt thereof,

wherein x is 1; m is 2,3, 4, 5,6, 7 or 8; y is H, OH or OMe; w is O or S; and each R_aIs that

In another aspect, compounds having the formula, or a pharmaceutically acceptable salt thereof,

wherein x is 1; m is 2,3, 4, 5,6, 7 or 8; y is H, OH or OMe; w is O or S; and each R_aIs that

In another aspect, the invention provides a pharmaceutical composition comprising at least one compound as described herein and a pharmaceutically acceptable carrier or diluent.

In yet another aspect, the present invention provides a method of treating or preventing a viral infection in a mammalian species in need thereof, the method comprising administering to the mammalian species a therapeutically effective amount of at least one compound described herein. In certain embodiments, the viral infection is an HIV infection. In certain other embodiments, the viral infection is an HBV infection. In yet another embodiment, the viral infection is an HCV infection. In yet another embodiment, the viral infection is an influenza a virus infection, a severe acute respiratory syndrome coronavirus infection, or a vaccinia virus infection.

In another aspect, the present invention provides a method of treating or preventing hepatitis c virus infection in a mammalian species in need thereof, the method comprising administering to the mammalian species a therapeutically effective amount of at least one compound described herein.

In another aspect, the present invention provides a method of inhibiting cyclophilin in a subject in need thereof, comprising administering to the subject an amount of at least one compound as described herein effective to inhibit cyclophilin.

In another aspect, the present invention provides a method of treating or preventing a cyclophilin-mediated disease in a mammalian species in need thereof, comprising administering to the mammalian species a therapeutically effective amount of at least one compound as described herein.

In another aspect, the present invention provides a method of treating or preventing a disease in a mammalian species in need thereof, the method comprising administering to the mammalian species a therapeutically effective amount of at least one compound as described herein, wherein the disease is selected from the group consisting of inflammation, respiratory inflammation, rheumatoid arthritis, and dry eye.

In yet another aspect, the invention provides a method of treating or preventing a disease in a mammalian species in need thereof, the method comprising administering to the mammalian species a therapeutically effective amount of at least one compound as described herein, wherein the disease is selected from the group consisting of neurodegenerative diseases, such as alzheimer's disease, parkinson's disease, huntington's disease, and ALS; traumatic brain injury; stroke; and ischemia reperfusion injury in brain, heart, kidney.

In a further aspect, the present invention provides a method of treating or preventing a disease in a mammalian species in need thereof, the method comprising administering to said mammalian species a therapeutically effective amount of at least one compound as described herein, wherein said disease is selected from cardiovascular disease, vascular stenosis, atherosclerosis, abdominal aortic aneurysm, cardiac hypertrophy, aortic rupture, pulmonary hypertension, myocarditis and myocardial fibrosis, and ischemic heart disease.

In yet another aspect, the invention provides a method of treating or preventing a disease or condition in a mammalian species in need thereof, the method comprising administering to the mammalian species a therapeutically effective amount of at least one compound as described herein, wherein the disease or condition is selected from cancer; obesity; diabetes mellitus; muscular dystrophy; lung and liver and kidney diseases and their protection; and alopecia.

In yet another aspect, the invention provides a method of treating or preventing a disease or condition in a mammalian species in need thereof, the method comprising administering to the mammalian species a therapeutically effective amount of at least one compound as described herein, wherein the disease or condition is selected from allergic conjunctivitis, atopic and vernal keratoconjunctivitis, atopic keratoconjunctivitis, anterior uveitis, behcet's disease, blepharitis, chronic ocular surface inflammation resulting from viral infection, corneal graft rejection, impaired corneal sensitivity due to surgery of the cornea or other ocular surface, meibomian gland disease, pterygium (ptyregia), ocular symptoms of graft versus host disease, ocular allergy, ocular cicatricial pemphigoid, Steven's syndrome, vernal keratoconjunctivitis, uveitis, herpes simplex keratitis, ocular rosacea, and meibomian spots.

Preparation method

In certain embodiments, compounds of formula (I) may be prepared by modifying cyclosporin a (cyclosporine a) at position 1 of MeBmt using similar methods for the synthesis of intermediates 3,4 and 5 as described in U.S. patent No. 9,200,038B2 (which is incorporated herein by reference) and U.S. patent application No. 2013/0190223a1 (which is incorporated herein by reference). ClAc in pyridine for cyclosporin A₂O and DMAP treatment followed by OsO in dioxane₄And NaIO₄Cleaving the double bond to give the corresponding aldehyde2And by wittig olefination with NaBH₄Reduction to give amines4Then coupled with AcOH to give amides5Which after hydrolysis gives the amides thereof6By a similar process as described in WO2012/051194a1 (which is incorporated herein by reference) and U.S. patent application No. 2013/0210704a1 (which is incorporated herein by reference), α -methylene is introduced at the sarcosine at position 3 of cyclosporine, giving it significant hydroxyl groupsIntermediates7When 1, 4-Michael addition is performed on methylene using a sulfur nucleophile, a methylene sulfur side chain with S-conformation can be formed on the sarcosine at position 3, and is a novel cyclosporin derivative. For example:

scheme 1

Other compounds disclosed herein can be prepared by using methods similar to those disclosed in scheme 1 and/or methods including any modifications or variations thereof known to those of ordinary skill in the art.

Recently, we published our studies on STG-175 (Gallay, P.A., et al, 2016, PLoS One11(4): e0152036.doi:10.1371/journal. bone.0152036, incorporated herein by reference). It has been demonstrated that cyclosporin has a CH in position 3 of sarcosine₂-S-CH₂CH₂CH₂CH₂OH side has high cyclophilin binding (EC for cyclophilin A)₅₀0.6nM) and has high anti-HCV activity (EC for polygenic HCV GT1 a-4 a)₅₀11.5-38.9 nM). In contrast to Sofosbuvir (Sofosbuvir), STG-175 still clears cells from HCV when their viral replication rebounds, since no viral replication rebound is observed after discontinuation of drug treatment. Even though the drug dose is 2.5 times lower than that of sofosbuvir. In addition, it has a higher resistance barrier than other cyclophilin inhibitors or Direct Antiviral Agents (DAAs). No cross-resistance was observed with DAA. Thus, from this finding, we believe that by modifying the CH at position 1 of MeBmt and position 3 of the sarcosine of cyclosporin₂-S-CH₂CH₂CH₂CH₂The OH side is a novel cyclosporin derivative, and a better derivative can be prepared.

Pharmaceutical composition

The present invention also provides a pharmaceutical composition comprising at least one compound as described herein, or a pharmaceutically acceptable salt or solvate thereof, and a pharmaceutically acceptable carrier.

The phrase "pharmaceutically acceptable carrier" as used herein refers to a pharmaceutically acceptable substance, composition or vehicle, such as a liquid or solid filler, diluent, excipient, solvent or encapsulating material, involved in carrying or transporting a primary medicament from one organ, or portion of the body, to another organ, or portion of the body. Each carrier must be "acceptable" in the sense of being compatible with the other ingredients of the formulation and not injurious to the patient. Some examples of substances that may serve as pharmaceutically acceptable carriers include: sugars such as lactose, glucose and sucrose; starches, such as corn starch and potato starch; cellulose and its derivatives, such as sodium carboxymethyl cellulose, ethyl cellulose and cellulose acetate; tragacanth powder; malt; gelatin; talc; excipients, such as cocoa butter and suppository waxes; oils such as peanut oil, cottonseed oil, safflower oil, sesame oil, olive oil, corn oil and soybean oil; glycols, such as butanediol; polyols such as glycerol, sorbitol, mannitol and polyethylene glycol; esters, such as ethyl oleate and ethyl laurate; agar; buffering agents such as magnesium hydroxide and aluminum hydroxide; alginic acid; pyrogen-free water; isotonic saline; ringer's solution; ethanol; phosphate buffer; and other non-toxic compatible substances used in medicaments.

As noted above, certain embodiments of the agents of the present invention may be provided in the form of pharmaceutically acceptable salts. In this regard, the term "pharmaceutically acceptable salts" refers to the relatively non-toxic, inorganic and organic acid addition salts of the compounds of the present invention. These salts may be prepared in situ during the final isolation and purification of the compounds of the invention, or formed by reacting the purified compounds of the invention in their free base form separately with a suitable organic or inorganic acid, and isolating the salt. Representative salts include hydrobromide, hydrochloride, sulfate, bisulfate, phosphate, nitrate, acetate, valerate, oleate, palmitate, stearate, laurate, benzoate, lactate, phosphate, tosylate, citrate, maleate, fumarate, succinate, tartrate, naphthenate, mesylate, glucoheptonate, lactobionate and laurylsulfonate salts, and the like. (see, e.g., Berge et al, (1977) "Pharmaceutical Salts", J.pharm.Sci.66: 1-19).

Pharmaceutically acceptable salts of the compounds of the invention include the conventional non-toxic salts or quaternary ammonium salts of the compounds, for example, from non-toxic organic or inorganic acids. For example, such conventional non-toxic salts include those derived from inorganic acids such as hydrochloric acid, hydrobromic acid, sulfuric acid, sulfamic acid, phosphoric acid, nitric acid, and the like; and salts prepared from organic acids such as acetic acid, butyric acid (butyric acid), succinic acid, glycolic acid, stearic acid, lactic acid, malic acid, tartaric acid, citric acid, ascorbic acid, palmitic acid, maleic acid, hydroxymaleic acid, phenylacetic acid, glutamic acid, benzoic acid, salicylic acid, sulfanilic acid, 2-acetoxybenzoic acid, fumaric acid, toluenesulfonic acid, methanesulfonic acid, ethanedisulfonic acid, oxalic acid, isothiocarboxylic acid (isothionic) and the like.

In other cases, the compounds of the invention may contain one or more acidic functional groups and, therefore, are capable of forming pharmaceutically acceptable salts with pharmaceutically acceptable bases. The term "pharmaceutically acceptable salts" in these instances refers to the relatively non-toxic inorganic and organic base addition salts of the compounds of the present invention. These salts can also be prepared in situ during the final isolation and purification of the compounds, or by reacting the purified compounds in their free acid form with a suitable base (such as a hydroxide, carbonate or bicarbonate of a pharmaceutically acceptable metal cation), ammonia or a pharmaceutically acceptable organic primary, secondary or tertiary amine, alone. Representative alkali or alkaline earth metal salts include lithium, sodium, potassium, calcium, magnesium, and aluminum salts and the like. Representative organic amines useful in the formation of base addition salts include ethylamine, diethylamine, ethylenediamine, ethanolamine, diethanolamine, piperazine and the like. (see, e.g., Berge et al, supra)

Wetting agents, emulsifiers and lubricants, such as sodium lauryl sulfate, magnesium stearate and polyethylene oxide-polybutylene oxide copolymers, as well as coloring, releasing, coating, sweetening, flavoring and perfuming agents, preservatives and antioxidants can also be present in the composition.

The formulations of the present invention include those suitable for oral, nasal, topical (including buccal and sublingual), rectal, vaginal and/or parenteral administration. The formulations may conveniently be presented in unit dosage form and may be prepared by any of the methods well known in the art of pharmacy. The amount of active ingredient that may be combined with the carrier materials to produce a single dosage form will vary depending upon the host treated and the particular mode of administration. The amount of active ingredient that can be combined with the carrier materials to produce a single dosage form will generally be that amount of the compound which produces a therapeutic effect. Typically, the amount is in the range of about 1% to about 99%, preferably about 5% to about 70%, most preferably about 10% to about 30% of the active ingredient, by 100%.

The methods of making these formulations or compositions include the step of admixing a compound of the present invention with a carrier and optionally one or more accessory ingredients. In general, the formulations are prepared by uniformly and intimately admixing the compounds of the invention with liquid carriers or finely divided solid carriers or both, and then, if necessary, shaping the product.

Formulations of the invention suitable for oral administration may be in the form of capsules, cachets, pills, tablets, lozenges (using a flavored basis, typically sucrose and acacia or tragacanth), powders, granules, or as a solution or suspension in an aqueous or non-aqueous liquid, or as an oil-in-water or water-in-oil liquid emulsion, or as an elixir or syrup, or as pastilles (using an inert base such as gelatin and glycerin, or sucrose and acacia) and/or mouthwashes and the like, each containing a predetermined amount of a compound of the invention as an active ingredient. The compounds of the invention may also be administered as a bolus, electuary or paste.

In solid dosage forms of the invention for oral administration (capsules, tablets, pills, dragees, powders, granules and the like), the active ingredient is mixed with one or more pharmaceutically acceptable carriers, such as sodium citrate or dicalcium phosphate, and/or any of the following: fillers or extenders, such as starch, lactose, sucrose, glucose, mannitol, and/or silicic acid; binders, such as, for example, carboxymethylcellulose, alginates, gelatin, polyvinylpyrrolidone, sucrose and/or acacia; humectants, such as glycerol; disintegrating agents, such as agar-agar, calcium carbonate, potato or tapioca starch, alginic acid, certain silicates, sodium carbonate, and sodium starch glycolate; solution retarding agents, such as paraffin; absorption promoters, such as quaternary ammonium compounds; wetting agents, such as, for example, cetyl alcohol, glycerol monostearate and polyethylene oxide-polybutylene oxide copolymers; absorbents such as kaolin and bentonite clays; lubricants, such as talc, calcium stearate, magnesium stearate, solid polyethylene glycols, sodium lauryl sulfate and mixtures thereof; and a colorant. In the case of capsules, tablets and pills, the pharmaceutical compositions may also comprise buffering agents. Solid compositions of a similar type may also be employed as fillers in soft and hard-filled gelatin capsules using excipients such as lactose or milk sugar as well as high molecular weight polyethylene glycols and the like.

Tablets may be prepared by compression or moulding, preferably together with one or more accessory ingredients. Compressed tablets may be prepared using binders (for example, gelatin or hydroxybutyl methylcellulose), lubricants, inert diluents, preservatives, disintegrating agents (for example, sodium starch glycolate or cross-linked sodium carboxymethyl cellulose), surface-active or dispersing agents. Molded tablets may be made by molding in a suitable machine a mixture of the powdered compound moistened with an inert liquid diluent.

Tablets and other solid dosage forms of the pharmaceutical compositions of the present invention, such as dragees, capsules, pills and granules, can optionally be scored or prepared with coatings and shells, such as enteric coatings and other coatings well known in the pharmaceutical art. They may also be formulated to provide slow or controlled release of the active ingredient therein using, for example, hydroxybutyl methylcellulose from various buttorions to provide the desired release profile, other polymer matrices, liposomes and/or microspheres. They may be sterilized, for example, by filtration through a bacterial-retaining filter or by incorporating sterilizing agents in the form of sterile solid compositions that can be dissolved in sterile water, or some other sterile injectable medium ready for use. These compositions may also optionally contain opacifying agents and may be of a composition that they release the active ingredient(s) only, or preferentially, in a certain portion of the gastrointestinal tract, optionally, in a delayed manner. Examples are embedding compositions, which may be used including polymeric substances and waxes. The active ingredient may also be in the form of microcapsules, if appropriate together with one or more of the above-mentioned excipients.

In addition to the active ingredient, the liquid dosage forms may contain inert diluents commonly used in the art such as, for example, water or other solvents, solubilizing agents and emulsifiers, such as ethyl alcohol, isobutyl alcohol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, butylene glycol, 1, 3-butylene glycol, oils (in particular, cottonseed, groundnut, corn, germ, olive, castor, and sesame oils), glycerol, tetrahydrofuryl alcohol, polyethylene glycols and fatty acid esters of sorbitan, and mixtures thereof.

In addition to inert diluents, the oral compositions can also include adjuvants such as wetting agents, emulsifying and suspending agents, sweetening, flavoring, coloring, perfuming and preservative agents.

Suspensions, in addition to the active compounds, may contain suspending agents as, for example, ethoxylated isostearyl alcohols, polyoxyethylene sorbitol and sorbitan esters, microcrystalline cellulose, aluminum metahydroxide, bentonite, agar-agar and tragacanth, and mixtures thereof.

Formulations of the pharmaceutical compositions of the present invention for rectal or vaginal administration may be presented as a suppository, which may be prepared by mixing one or more compounds of the present invention with one or more suitable non-irritating excipients or carriers comprising, for example, cocoa butter, polyethylene glycol, a suppository wax or a salicylate, and which is solid at room temperature but liquid at body temperature and, therefore, will melt in the rectum or vaginal cavity and release the active agent of the present invention.

Formulations of the invention suitable for vaginal administration also include pessaries, tampons, creams, gels, pastes, foams or spray formulations containing such carriers as are known in the art to be suitable.

Dosage forms for topical or transdermal administration of the compounds of the present invention include powders, sprays, ointments, pastes, creams, lotions, gels, solutions, patches and inhalants. The active compound may be mixed under sterile conditions with a pharmaceutically acceptable carrier, and with any preservatives, buffers, or propellants (bunellant) that may be required.

Ointments, pastes, creams and gels may contain, in addition to an active compound of the invention, excipients, for example animal and vegetable fats, oils, waxes, paraffins, starch, tragacanth, cellulose derivatives, polyethylene glycols, silicones, bentonites, silicic acid, talc and zinc oxide, or mixtures thereof.

Powders and sprays can contain, in addition to a compound of the invention, excipients such as lactose, talc, silicic acid, aluminum hydroxide, calcium silicates and polyamide powder or mixtures of these substances. Sprays can additionally contain customary propellants (butellant), such as chlorofluorohydrocarbons and volatile unsubstituted hydrocarbons, such as butane and butane.

Transdermal patches have the additional advantage of providing controlled delivery of the compounds of the present invention to the body. Such dosage forms may be prepared by dissolving or dispersing the agent in a buffered medium. Absorption enhancers may also be used to increase the flux of the agents of the invention through the skin. The rate of such flux can be controlled by providing a rate controlling membrane or dispersing the compound in a polymer matrix or gel.

Ophthalmic formulations, ophthalmic ointments, powders, solutions, and the like are also contemplated within the scope of the present invention.

Pharmaceutical compositions of the invention suitable for parenteral administration comprise one or more compounds of the invention in combination with one or more pharmaceutically acceptable sterile isotonic aqueous or non-aqueous solutions, dispersions, suspensions or emulsions, or sterile powders which may be reconstituted into sterile injectable solutions or dispersions prior to use, which may contain antioxidants, buffers, bacteriostats, solutes which render the formulation isotonic with the blood of the intended recipient, or suspending or thickening agents.

In some cases, to prolong the effect of a drug, it is desirable to slow the absorption of the drug from subcutaneous or intramuscular injection. This can be achieved by using a liquid suspension of crystalline or amorphous material which is poorly water soluble. The rate of absorption of the drug then depends on its rate of dissolution, which in turn may depend on crystal size and crystal form. Alternatively, delayed absorption of a parenterally administered drug form is achieved by dissolving or suspending the drug in an oily vehicle. One strategy for long acting injections includes the use of polyethylene oxide-polybutylene oxide copolymers where the vehicle is fluid at room temperature and solidifies at body temperature.

Injectable depot forms are made by forming a microcapsule matrix of the compounds of the present invention in a biodegradable polymer such as polylactide-polyglycolide. Depending on the ratio of drug to polymer, and the nature of the particular polymer used, the rate of drug release can be controlled. Examples of other biodegradable polymers include poly (orthoesters) and poly (anhydrides). Depot injectable formulations can also be prepared by entrapping the drug in liposomes or microemulsions which are compatible with body tissues.

When the compounds of the present invention are administered as medicaments to humans and animals, they may be administered alone or as a pharmaceutical composition containing, for example, 0.1% to 99.5% (more preferably, 0.5% to 90%) of the active ingredient in combination with a pharmaceutically acceptable carrier.

The compounds and pharmaceutical compositions of the present invention may be used in combination therapy, that is, the compounds and pharmaceutical compositions may be administered simultaneously, prior to, or after one or more other desired therapeutic agents or procedures. The particular combination of therapies (therapeutic agents or procedures) used in a combination regimen will take into account the compatibility of the therapeutic agents and/or procedures required and the therapeutic effect desired to be achieved. It will also be appreciated that the therapies employed may achieve the desired effect against the same condition (e.g., the compounds of the present invention may be administered simultaneously with another anti-HCV drug), or they may achieve different effects (e.g., control of any adverse effects).

The compounds of the present invention may be administered intravenously, intramuscularly, intraperitoneally, subcutaneously, topically, orally, or by other acceptable means. The compounds are useful for treating arthritic disorders in mammals (i.e., humans, livestock and domesticated animals), birds, lizards and any other organism that can tolerate the compounds.

The invention also provides a pharmaceutical pack or kit comprising one or more containers filled with one or more of the ingredients of the pharmaceutical composition of the invention. Optionally associated with such container(s) may be a notice in the form prescribed by a governmental agency regulating the manufacture, use or sale of pharmaceuticals or biologicals, which notice reflects approval by the agency for manufacture, use or sale for human administration.

Equivalents of

The following representative examples are set forth to aid in the description of the invention and are not intended to, nor should they be construed to, limit the scope of the invention. Indeed, various modifications of the invention and many of its further embodiments, in addition to those shown and described herein, will become apparent to those skilled in the art from the entire disclosure of this document, including the following and the examples referenced in the scientific and patent literature cited herein. It should also be understood that the contents of these cited references are incorporated herein by reference to help explain the state of the art. The following examples contain important additional information, exemplification and guidance which are suitable for the practice of this invention in its various embodiments and equivalents.

Examples

Example 1

[ α -methylene-Sar]-3-Cyclosporin

To [ α -hydroxymethyl-Sar ]]To a solution of (246mg, 0.20mmol) of (E) -3-cyclosporin in tetrahydrofuran (15ml) were added sodium hydride (120mg, 60% in mineral oil, 3mmol) and diethyl chlorophosphate (412mg, 2.40 mmol). The resulting mixture was stirred at room temperature overnight. After disappearance of the starting material, methanol (10ml) and potassium tert-butoxide (33mg, 26.00mmol) were added. The mixture was stirred at room temperature for 3 hours, and the solvent was removed under reduced pressure. The residue was dissolved in dichloromethane (50 ml). The dichloromethane layer was washed with brine, dried over magnesium sulfate and evaporated under reduced pressure. The residue was purified by flash chromatography using dichloromethane/methanol as eluent to give 205mg of the product [ formula: c₆₃H₁₁₁N₁₁O₁₂(ii) a Accurate quality: 1213.84, respectively; MS (M/z) 1214.59(M +1)⁺(ii) a HPLC RT: 17.47 min. (C8 reverse phase column: 250 mm; acetonitrile/water (0.05% trifluoroacetic acid), operating temperature: 64 ℃ C.; detector: 210nm)]。

Example 2

[ (3R,4R) -3-hydroxy-4-methyl-6- (4-methoxycarbonylbenzyl) -N-Menle]-1- [ (S) - (4-Hydroxybutane) Thio) methyl-Sar]-3-Cyclosporin

[ 3-acetyl-MeBmt ] -1-Cyclosporin

To a dry flask was added cyclosporin (12.00g, 9.98mmol), N-dimethylaminopyridine (0.12g, 0.10mmol), anhydrous pyridine (120ml) and acetic anhydride (54ml, 0.54mol) under nitrogen. After stirring at room temperature overnight, the mixture was poured into ice water (600ml) and stirred until the ice melted. Ethyl acetate (100ml) was added and the mixture was separated. The ethyl acetate layer was washed with 1.0N hydrochloric acid solution (100 ml. times.2), saturated sodium bicarbonate solution (100ml), brine (100ml), dried over magnesium sulfate and evaporated under reduced pressure. The residue was purified by chromatography (hexane/acetone) to give 11.40g of pure [ 3-acetyl-MeBmt]-1-cyclosporin [ formula: c₆₄H₁₁₃N₁₁O₁₃(ii) a Accurate quality: 1243.85, respectively; MS (M/z) 1244.53(M +1)⁺,1266.70(M+Na)⁺(ii) a HPLC RT: 18.02 min. (C8 reverse phase column: 250 mm; acetonitrile/water (0.05% trifluoroacetic acid); operating temperature: 64 ℃ C.; detector: 210nm)]。

[ (3R,4R) -3-acetoxy-4-methyl-6-oxo-N-Menle ] -1-Cyclosporin

To [ 3-acetyl-MeBmt]To a solution of-1-cyclosporin (10.00g, 8.04mmol) in dioxane (200ml) was added water (20ml), osmium (VIII) oxide solution (15.74mM, 51ml, 0.80mmol) and sodium metaperiodate (6.88g, 32.16 mmol). The reaction mixture was stirred at room temperature for 5 hours. Ethyl acetate (100ml) and brine (100ml) were added and the mixture was separated. The organic layer was washed with saturated sodium bicarbonate solution, brine, dried over magnesium sulfate and evaporated under reduced pressure. The residue was purified by chromatography (hexane/acetone) to give 5.10g of pure [ (3R,4R) -3-acetoxy-4-methyl-6-oxo-N-Menle]-1-cyclosporin [ formula: c₆₂H₁₀₉N₁₁O₁₄(ii) a Accurate quality: 1231.82, respectively; MS (M/z) 1232.75(M +1)⁺,1254.77(M+Na)⁺(ii) a HPLC RT: 15.74 min. (C8 reverse phase column: 250 mm; acetonitrile/water (0.05% trifluoroacetic acid); operating temperature: 64 ℃ C.; detector: 210nm)]。

[ (3R,4R) -3-acetoxy-4-methyl-6- (4-methoxycarbonylphenyl) methylene-N-Menle ] -1-cyclosporin

To a solution of (4-methoxycarbonylbenzyl) triphenyl phosphonium bromide (4.00g, 8.14mmol) in anhydrous tetrahydrofuran (120ml) was added sodium bis (trimethylsilyl) amide (1.0M in THF, 10ml, 10.00mmol) under nitrogen. The reaction mixture was stirred at room temperature for one hour and cooled to-40 ℃. Adding [ (3R,4R) -3-acetoxy-4-methyl-6-oxo-N-Menle]-1-Cyclosporin (5.00g, 4.05mmol) in anhydrous tetrahydrofuran (25 ml). The mixture was stirred at-30 ℃ for a further 2 hours and saturated ammonium chloride solution (20ml) was added to quench the reaction. Most of the tetrahydrofuran was evaporated under reduced pressure. Ethyl acetate (150ml) and brine (50ml) were added and the mixture was separated. The organic layer was dried over magnesium sulfate and evaporated under reduced pressure. The residue was purified by chromatography (dichloromethane/methanol) to give 2.00g of pure [ (3R,4R) -3-acetoxy-4-methyl-6- (4-methoxycarbonylphenyl) methylene-N-Menle]-1-cyclosporin [ formula: c₇₁H₁₁₇N₁₁O₁₅(ii) a Accurate quality: 1363.87, respectively; MS (M/z) 1364.61(M +1)⁺HPLC RT:17.89 min. (C8 reverse phase column: 250 mm; acetonitrile/water (0.05% trifluoroacetic acid); operating temperature: 64 ℃ C.; detector: 210nm)]。

[ (3R,4R) -3-acetoxy-4-methyl-6- (4-methoxycarbonylbenzyl) -N-Menle ] -1-Cyclosporin

To [ (3)R,4R) -3-acetoxy-4-methyl-6- (4-methoxycarbonylphenyl) methylene-N-Menle]To a solution of-1-cyclosporin (2.00g, 1.46mmol) in methanol (50ml) was added palladium (10 wt% on carbon, 20mg) and acetic acid (5 drops). The mixture was stirred at room temperature under hydrogen for 2 hours. The mixture was filtered and the filtrate was evaporated under reduced pressure to give crude [ (3R,4R) -3-acetoxy-4-methyl-6- (4-methoxycarbonylbenzyl) -N-Menle]-1-cyclosporin [ formula: c₇₁H₁₁₉N₁₁O₁₅(ii) a Accurate quality: 1365.89, respectively; MS (M/z) 1366.73(M +1)⁺(ii) a HPLC RT: 18.02 min. (C8 reverse phase column: 250 mm; acetonitrile/water (0.05% trifluoroacetic acid); operating temperature: 64 ℃ C.; detector: 210nm)]。

[ (3R,4R) -3-hydroxy-4-methyl-6- (4-carboxybenzyl) -N-Menle ] -1-cyclosporin

Reacting [ (3R,4R) -3-acetoxy-4-methyl-6- (4-methoxycarbonylbenzyl) -N-Menle]-1-Cyclosporin (0.25g, 0.18mmol) was dissolved in methanol (4 ml). Water (2ml) and tetramethylammonium hydroxide pentahydrate (70mg) were added. The mixture was stirred at room temperature overnight. Most of the methanol is then evaporated. The pH of the mixture was adjusted to 5 with acetic acid. Ethyl acetate (50ml) and brine (10ml) were added and the mixture was separated. The organic layer was dried over magnesium sulfate and evaporated under reduced pressure to give the crude [ (3R,4R) -3-hydroxy-4-methyl-6- (4-carboxybenzyl) -N-Menle]-1-cyclosporin [ formula: c₆₈H₁₁₅N₁₁O₁₄(ii) a Accurate quality: 1309.86, respectively; MS (M/z) 1310.61(M +1)⁺(ii) a HPLC RT: 14.36 min. (C8 reverse phase column: 250 mm; acetonitrile/water (0.05% trifluoroacetic acid); operating temperature: 64 ℃ C.; detector: 210nm)]。

[ (3R,4R) -3-hydroxy-4-methyl-6- (4-carboxybenzyl) -N-Menle ] -1- [ α -methylene-Sar ] -3-cyclosporin

N-butyllithium (2.65M, 7.7ml, 20.41mmol) was added to a solution of diisopropylamine (2.05g, 20.30mmol) in tetrahydrofuran (50ml) at-78 ℃ under nitrogen. After stirring the reaction mixture for 1 hour, [ (3R,4R) -3-hydroxy-4-methyl-6- (4-carboxybenzyl) -N-Menle was added]-1-Cyclosporin (2.40g, 1.20mmol) in tetrahydrofuran (10 ml). The mixture was stirred at-78 ℃ for three hours. After bubbling carbon dioxide gas into the reaction mixture for 15 minutes, the mixture was stirred at-78 ℃ for another 1 hour. The cooling bath was then removed and the reaction mixture was allowed to warm to room temperature to allow unreacted carbon dioxide to vent. The mixture was again cooled to-78 ℃ and chloromethyl chloroformate (3.00ml) was added. The mixture was stirred and allowed to warm to room temperature overnight. Brine (5ml) was added to quench the reaction. Most of the tetrahydrofuran was removed under reduced pressure. Ethyl acetate (80ml) and brine (50ml) were added and the mixture was separated. The organic layer was dried over magnesium sulfate and evaporated under reduced pressure to give 1.50g of a crude product [ (3R,4R) -3-hydroxy-4-methyl-6- (4-carboxybenzyl) -N-Menle]-1- [ α -methylene-Sar]-3-cyclosporin [ formula: c₆₉H₁₁₅N₁₁O₁₄(ii) a Accurate quality: 1321.86, respectively; MS (M/z) 1322.55(M +1)⁺(ii) a HPLC RT: 16.13 min. (C8 reverse phase column: 250 mm; acetonitrile/water (0.05% trifluoroacetic acid); operating temperature: 64 ℃ C.; detector: 210nm)]。

[ (3R,4R) -3-hydroxy-4-methyl-6- (4-methoxycarbonylbenzyl) -N-Menle ] -1- [ α -methylene-Sar ] -3-cyclosporin

To [ (3R,4R) -3-hydroxy-4-methyl-6- (4-carboxybenzyl) -N-Menle]-1- [ α -methylene-Sar]To a solution of (1.32g, 1.00mmol) of (E) -3-cyclosporin in N, N-dimethylformamide (15ml) were added potassium carbonate (0.38g, 2.75mmol) and methyl iodide (0.50g, 3.52 mmol). The mixture was stirred at room temperature for three hours. Ethyl acetate (100ml) and water (50ml) were added to the solution, and the mixture was separated. Drying with magnesium sulfateThe organic layer was evaporated under reduced pressure. The residue was purified by chromatography (dichloromethane/methanol) to give 1.60g of pure [ (3R,4R) -3-hydroxy-4-methyl-6- (4-methoxycarbonylbenzyl) -N-Menle]-1- [ α -methylene-Sar]-3-cyclosporin [ formula: c₇₀H₁₁₇N₁₁O₁₄(ii) a Accurate quality: 1335.88, respectively; MS (M/z) 1336.64(M +1)⁺(ii) a HPLC RT: 18.03 min. (C8 reverse phase column: 250 mm; acetonitrile/water (0.05% trifluoroacetic acid); operating temperature: 64 ℃ C.; detector: 210nm)]。

[ (3R,4R) -3-hydroxy-4-methyl-6- (4-methoxycarbonylbenzyl) -N-Menle ] -1- [ (S) - (4-hydroxybutylthio) methyl-Sar ] -3-cyclosporin

To [ (3R,4R) -3-hydroxy-4-methyl-6- (4-methoxycarbonylbenzyl) -N-Menle]-1- [ α -methylene-Sar]To a solution of (0.60g, 0.45mmol) of (E) -3-cyclosporin in methanol (25ml) were added (0.45g, 4.25mmol) of 4-mercapto-1-butanol and (0.10g, 4.25mmol) of lithium hydroxide. The reaction mixture was stirred at room temperature overnight. Most of the methanol was evaporated under reduced pressure. Ethyl acetate (60ml) and brine (30ml) were added and the mixture was separated. The organic layer was dried over magnesium sulfate and evaporated under reduced pressure. The residue was purified by chromatography (dichloromethane/methanol) to give pure [ (3R,4R) -3-hydroxy-4-methyl-6- (4-methoxycarbonylbenzyl) -N-Menle]-1- [ (S) - (4-hydroxybutylthio) methyl-Sar]-3-cyclosporin [ formula: c₇₄H₁₂₇N₁₁O₁₅S; accurate quality: 1441.92, respectively; MS (M/z) 1422.65(M +1)⁺(ii) a HPLC RT: 16.79 min. (C8 reverse phase column: 250 mm; acetonitrile/water (0.05% trifluoroacetic acid); operating temperature: 64 ℃ C.; detector: 210nm)]。

Example 3

[ (3R,4R) -3-hydroxy-4-methyl-6- (4-carboxybenzyl) -N-Menle]-1- [ (S) - (4-hydroxybutylthio) methyl-Sar]-3-Cyclosporin

To [ (3R,4R) -3-hydroxy-4-methyl-6- (4-methoxycarbonylbenzyl) -N-Menle]-1- [ (S) - (4-hydroxybutylthio) methyl-Sar]To a solution of-3-cyclosporin (85mg, 0.06mmol) in methanol (5ml) was added a solution of lithium hydroxide (15mg) in water (5 ml). The reaction mixture was stirred at room temperature overnight. Most of the methanol was evaporated under reduced pressure. The pH of the mixture was adjusted to 6 with 1.0N hydrochloric acid. Ethyl acetate (60ml) and brine (10ml) were added to the solution, and the mixture was separated. The organic layer was dried over magnesium sulfate and evaporated under reduced pressure. Purifying the residue to obtain pure [ (3R,4R) -3-hydroxy-4-methyl-6- (4-carboxybenzyl) -N-Menle]-1- [ (S) - (4-hydroxybutylthio) methyl-Sar]-3-cyclosporin [ formula: c₇₃H₁₂₅N₁₁O₁₅S; accurate quality: 1427.91, respectively; MS (M/z) 1428.63(M +1)⁺(ii) a HPLC RT: 14.37 min. (C8 reverse phase column: 250 mm; acetonitrile/water (0.05% trifluoroacetic acid); operating temperature: 64 ℃ C.; detector: 210nm)]。

Example 4

[ (3R,4R) -3-hydroxy-4-methyl-6- (3-amino) benzyl-N-Menle]-1- [ (S) - (4-hydroxybutylthio) methyl-Sar]-3-Cyclosporin

(3-nitrobenzyl) triphenylphosphine bromide

3-Nitrobenzylbromide (5.00g, 23.15mmol) and triphenylphosphine (6.06g, 23.15mmol) were added to toluene (60 ml). The mixture was stirred and heated to reflux overnight. After cooling to room temperature, the precipitate was filtered off, washed with toluene and hexane and dried in vacuo to yield 10.50g of product [ formula: c₂₅H₂₁BrNO₂P; accurate quality: 477.05, respectively; MS (m/z):398.45(M+1)⁺-Br]。

[ (3R,4R) -3-acetoxy-4-methyl-6- (3-nitro-phenylmethylene) -N-Menle ] -1-cyclosporin

To a solution of (3-nitrobenzyl) triphenyl phosphine bromide (2.40g, 5.03mmol) in anhydrous tetrahydrofuran (90ml) was added sodium bis (trimethylsilyl) amide (1.0M in THF, 6ml, 6.00mmol) under nitrogen. The reaction mixture was stirred at room temperature for one hour and cooled to-40 ℃. Adding [ (3R,4R) -3-acetoxy-4-methyl-6-oxo-N-Menle]-1-Cyclosporin (3.69g, 3.00mmol) in anhydrous tetrahydrofuran (25 ml). The mixture was stirred at-30 ℃ for a further 2 hours. Saturated ammonium chloride solution (20ml) was then added to quench the reaction. Most of the tetrahydrofuran was evaporated under reduced pressure. Ethyl acetate (150ml) and brine (50ml) were added and the mixture was separated. The organic layer was dried over magnesium sulfate and evaporated under reduced pressure. The residue was purified by chromatography (dichloromethane/methanol) to give 1.60g of pure [ (3R,4R) -3-acetoxy-4-methyl-6- (3-nitro-phenylmethylene) N-Menle]-1-cyclosporin [ formula: c₆₉H₁₁₄N₁₂O₁₅(ii) a Accurate quality: 1350.85, respectively; MS (M/z) 1351.52(M +1)⁺]。

[ (3R,4R) -3-acetoxy-4-methyl-6- (3-amino) benzyl-N-Menle ] -1-cyclosporin

To [ (3R,4R) -3-acetoxy-4-methyl-6- (3-nitro-phenylmethylene) -N-Menle under nitrogen]To a solution of (3.00g, 2.22mmol) of (E) -1-cyclosporin in methanol (50ml) was added nickel (II) chloride hexahydrate (0.19g, 0.81 mmol). The reaction mixture was placed in an ice-water bath. Sodium borohydride (1.80g, 47.33mmol) was added in four portions over a two hour period. After stirring the mixture at 0 ℃ for a further 2 hours,water (10ml) was added. Most of the methanol was evaporated under reduced pressure. Ethyl acetate (50ml) and saturated sodium bicarbonate solution (50ml) were added to the solution, and the mixture was separated. The organic layer was washed with brine, dried over magnesium sulfate and evaporated under reduced pressure. The residue was dissolved in methanol (30 ml). Palladium (10 wt% on carbon, 20mg) and acetic acid (5 drops) were added. The mixture was stirred at room temperature under hydrogen for 2 hours. The mixture was filtered and the filtrate was evaporated under reduced pressure to give crude [ (3R,4R) -3-acetoxy-4-methyl-6- (3-amino) benzyl-N-Menle]-1-cyclosporin [ formula: c₆₉H₁₁₈N₁₂O₁₃(ii) a Accurate quality: 1322.89, respectively; MS (M/z) 1323.73(M +1)⁺]。

[ (3R,4R) -3-hydroxy-4-methyl-6- (3-amino) benzyl-N-Menle ] -1-cyclosporin

Mixing [ (3R,4R) -3-acetoxyl-4-methyl-6- (3-amino) benzyl-N-Menle]-1-Cyclosporin (2.60g, 1.56mmol) was dissolved in methanol (70 ml). Water (35ml) and tetramethylammonium hydroxide pentahydrate (2.00g, 11.01mmol) were added. The mixture was stirred at room temperature for eight hours. Most of the methanol was evaporated. Ethyl acetate (100ml) and brine (100ml) were added and the mixture was separated. The organic layer was dried over magnesium sulfate and evaporated under reduced pressure. The residue was purified by chromatography (dichloromethane/methanol) to give 1.90g of pure [ (3R,4R) -3-hydroxy-4-methyl-6- (3-aminophenyl) methyl-N-Menle]-1-cyclosporin [ formula: c₆₇H₁₁₆N₁₂O₁₂(ii) a Accurate quality: 1280.88, respectively; MS (M/z) 1281.64(M +1)⁺]。

[ (3R,4R) -3-hydroxy-4-methyl-6- (3- (tert-butoxycarbonyl) amino) benzyl-N-Menle ] -1-cyclosporin

The [ (3R,4R) -3-hydroxy-4-methyl-6- (3-amino)) benzyl-N-Menle]-1-Cyclosporin (0.80g, 0.62mmol) was dissolved in tetrahydrofuran (30 ml). Di-tert-butyl dicarbonate (0.37g, 1.70mmol) was added. The mixture was stirred at room temperature for two days. Most of the tetrahydrofuran was evaporated under reduced pressure. Ethyl acetate (30ml) and brine (30ml) were added and the mixture was separated. The organic layer was dried over magnesium sulfate and evaporated under reduced pressure. The residue was purified by chromatography to give 750mg of pure [ (3R,4R) -3-hydroxy-4-methyl-6- (3- (tert-butoxycarbonyl) amino) benzyl-N-Menle]-1-cyclosporin [ formula: c₇₂H₁₂₄N₁₂O₁₄(ii) a Accurate quality: 1380.94, respectively; MS (M/z) 1381.65(M +1)⁺]。

[ (3R,4R) -3-hydroxy-4-methyl-6- (3- (tert-butoxycarbonyl) amino) benzyl-N-Menle ] -1- [ α -methylene-Sar ] -3-cyclosporin

N-butyllithium (2.6.5M, 2.70ml, 7.16mmol) was added to a solution of diisopropylamine (0.71g, 7.15mmol) in tetrahydrofuran (50ml) at-78 ℃ under nitrogen. After stirring the reaction mixture for 1 hour, [ (3R,4R) -3-hydroxy-4-methyl-6- (3- (tert-butoxycarbonyl) amino) benzyl-N-Menle was added]-1-Cyclosporin (0.90g, 0.65mmol) in tetrahydrofuran (2 ml). The mixture was stirred at-78 ℃ for three hours. After bubbling carbon dioxide gas into the reaction mixture for 15 minutes, the mixture was stirred at-78 ℃ for another 1 hour. The cooling bath was then removed and the reaction mixture was allowed to warm to room temperature to allow unreacted carbon dioxide to vent. The mixture was cooled to-78 ℃ and chloromethyl chloroformate (1.20ml) was added. The mixture was stirred and allowed to warm to room temperature overnight. Brine (5ml) was added to quench the reaction. Most of the tetrahydrofuran was removed under reduced pressure. Ethyl acetate (50ml) and brine (50ml) were added and the mixture was separated. The organic layer was dried over magnesium sulfate and evaporated under reduced pressure. The residue was purified by chromatography (hexane/acetone) to give 0.45g of pure [ (3R,4R) -3-hydroxy-4-methyl-6- (3- (tert-butoxycarbonyl) amino) benzyl-N-Menle]-1- [ α -methylene-Sar]-3-cyclosporin [ formula: c₇₃H₁₂₄N₁₂O₁₄(ii) a Accurate quality: 1392.94, respectively; MS (M/z) 1393.88(M +1)⁺]。

[ (3R,4R) -3-hydroxy-4-methyl-6- (3-amino) benzyl-N-Menle ] -1- [ α -methylene-Sar ] -3-cyclosporin

Reacting [ (3R,4R) -3-hydroxy-4-methyl-6- (3- (tert-butoxycarbonyl) amino) benzyl-N-Menle]-1- [ α -methylene-Sar]-3-Cyclosporin (0.80g, 0.57mmol) was dissolved in dichloromethane (15ml) and placed in an ice-water bath. Trifluoroacetic acid (7ml) was added. The mixture was stirred at room temperature for one hour. Another portion of dichloromethane (50ml) was added. The mixture was washed with brine (30ml) and saturated sodium bicarbonate solution (30ml), dried over magnesium sulphate and evaporated under reduced pressure. The residue was purified by chromatography (dichloromethane/methanol) to give 0.20g of pure [ (3R,4R) -3-hydroxy-4-methyl-6- (3-amino) benzyl-N-Menle]-1- [ α -methylene-Sar]-3-cyclosporin [ formula: c₆₈H₁₁₆N₁₂O₁₂(ii) a Accurate quality: 1292.88, respectively; MS (M/z) 1293.68(M +1)⁺]。

[ (3R,4R) -3-hydroxy-4-methyl-6- (3-amino) benzyl-N-Menle ] -1- [ (S) - (4-hydroxybutylthio) methyl-Sar ] -cyclosporin

To [ (3R,4R) -3-hydroxy-4-methyl-6- (3-amino) benzyl-N-Menle]-1- [ α -methylene-Sar]To a solution of (0.20g, 0.15mmol) of (E) -3-cyclosporin in methanol (10ml) were added (0.07g, 0.66mmol) of 4-mercapto-1-butanol and (19mg, 0.79mmol) of lithium hydroxide. The reaction mixture was stirred at room temperature overnight. Most of the methanol was evaporated under reduced pressure. Ethyl acetate (30ml) and brine (30ml) were added and the mixture was separated. The organic layer was dried over magnesium sulfate and evaporated under reduced pressure. The residue was purified by chromatography (dichloromethane/methanol)To obtain pure [ (3R,4R) -3-hydroxy-4-methyl-6- (3-amino) benzyl-N-Menle]-1- [ (S) - (4-hydroxybutylthio) methyl-Sar]-3-cyclosporin [ formula: c₇₂H₁₂₆N₁₂O₁₃S; accurate quality: 1398.93, respectively; MS (M/z) 1399.62(M +1)⁺(ii) a HPLC RT: 11.33 min. (C8 reverse phase column: 250 mm; acetonitrile/water (0.05% trifluoroacetic acid); operating temperature: 64 ℃ C.; detector: 210nm)]。

Example 5

[ (3R,4R) -3-hydroxy-4-methyl-6- (3- (N, N-diisopropylamino) carbonyl) benzyl-N-Menle]-1- [ (S) - (4-hydroxybutylthio) methyl-Sar]-3-Cyclosporin

(3-Methoxycarbonylbenzyl) triphenylphosphine bromide

Methyl (3-bromomethyl) benzoate (6.00g, 26.20mmol) and triphenylphosphine (6.86g, 26.18mmol) were added to toluene (60 ml). The mixture was stirred and heated to reflux overnight. After cooling to room temperature, the precipitate was filtered off, washed with toluene and hexane and dried in vacuo to give 11.05g of product. [ molecular formula: c₂₇H₂₄BrO₂P; accurate quality: 490.07, respectively; MS (M/z) 411.25(M +1)⁺-Br]。

[ (3R,4R) -3-acetoxy-4-methyl-6- (3-methoxycarbonylphenyl) methylene-N-Menle ] -1-cyclosporin

Under the condition of nitrogen, the nitrogen is added,to a mixture of (3-methoxycarbonylbenzyl) triphenyl phosphonium bromide (4.00g, 8.14mmol) in anhydrous tetrahydrofuran (120ml) was added sodium bis (trimethylsilyl) amide (1.0M in THF, 10ml, 10.00 mmol). The reaction mixture was stirred at room temperature for one hour and cooled to-40 ℃. Adding [ (3R,4R) -3-acetoxy-4-methyl-6-oxo-N-Menle]-1-Cyclosporin (5.00g, 4.05mmol) in anhydrous tetrahydrofuran (25 ml). The mixture was stirred at-30 ℃ for a further 2 hours. Saturated ammonium chloride solution (20ml) was then added to quench the reaction. Most of the tetrahydrofuran was evaporated under reduced pressure. Ethyl acetate (150ml) and brine (50ml) were added and the mixture was separated. The organic layer was dried over magnesium sulfate and evaporated under reduced pressure. The residue was purified by chromatography (dichloromethane/methanol) to give 2.00g of pure [ (3R,4R) -3-acetoxy-4-methyl-6- (3-methoxycarbonylphenyl) methylene-N-Menle]-1-cyclosporin [ formula: c₇₁H₁₁₇N₁₁O₁₅(ii) a Accurate quality: 1363.87, respectively; MS (M/z) 1364.61(M +1)⁺]。

[ (3R,4R) -3-acetoxy-4-methyl-6- (3-methoxycarbonyl) benzyl-N-Menle ] -1-cyclosporin

To [ (3R,4R) -3-acetoxy-4-methyl-6- (3-methoxycarbonylphenyl) methylene-N-Menle]To a solution of-1-cyclosporin (2.00g, 1.46mmol) in methanol (50ml) was added palladium (10 wt% on carbon, 20mg) and acetic acid (5 drops). The mixture was stirred at room temperature under hydrogen for 2 hours. The mixture was filtered and the filtrate was evaporated under reduced pressure to give crude [ (3R,4R) -3-acetoxy-4-methyl-6- (3-methoxycarbonyl) benzyl-N-Menle]-1-cyclosporin [ formula: c₇₁H₁₁₉N₁₁O₁₅(ii) a Accurate quality: 1365.89, respectively; MS (M/z) 1366.73(M +1)⁺]。

[ (3R,4R) -3-hydroxy-4-methyl-6- (3-carboxy) benzyl-N-Menle ] -1-cyclosporin

The [ (3R,4R) -3-acetoxyl-4-methyl-6- (3-methoxycarbonyl) benzyl-N-Menle]-1-Cyclosporin (0.25g, 0.18mmol) was dissolved in methanol (4 ml). Water (2ml) and tetramethylammonium hydroxide pentahydrate (70mg) were added. The mixture was stirred at room temperature overnight. Most of the methanol was evaporated. The pH of the mixture was adjusted to 6 with acetic acid. Ethyl acetate (50ml) and brine (10ml) were added and the mixture was separated. The organic layer was dried over magnesium sulfate and evaporated under reduced pressure. The residue was used in the next step without further purification. [ (3R,4R) -3-hydroxy-4-methyl-6- (3-carboxy) benzyl-N-Menle]-1-cyclosporin [ formula: c₆₈H₁₁₅N₁₁O₁₄(ii) a Accurate quality: 1309.86, respectively; MS (M/z) 1310.61(M +1)⁺]。

[ (3R,4R) -3-hydroxy-4-methyl-6- (3-methoxycarbonyl) benzyl-N-Menle ] -1-cyclosporin

To [ (3R,4R) -3-hydroxy-4-methyl-6- (3-carboxy) benzyl-N-Menle]To a solution of (1.80g, 1.37mmol) of (E) -1-cyclosporin in N, N-dimethylformamide (25ml) were added potassium carbonate (0.38g, 2.75mmol) and methyl iodide (0.50g, 3.52 mmol). The mixture was stirred at room temperature for three hours. Ethyl acetate (100ml) and water (50ml) were added to the solution, and the mixture was separated. The organic layer was dried over magnesium sulfate and evaporated under reduced pressure. The residue was purified by chromatography (dichloromethane/methanol) to give 1.60g of pure [ (3R,4R) -3-hydroxy-4-methyl-6- (3-methoxycarbonyl) benzyl-N-Menle]-1-cyclosporin [ formula: c₆₉H₁₁₇N₁₁O₁₄(ii) a Accurate quality: 1323.88, respectively; MS (M/z) 1324.64(M +1)⁺]。

[ (3R,4R) -3-hydroxy-4-methyl-6- ((3-N, N-diisopropylaminocarbonyl)) benzyl-N-Menle ] -1- [ α -methylene-Sar ] -3-cyclosporin

N-butyllithium (2.65M, 5ml, 13.33mmol) was added to a solution of diisopropylamine (1.37g, 13.30mmol) in tetrahydrofuran (50ml) at-78 ℃ under nitrogen. After stirring the reaction mixture for 1 hour, [ (3R,4R) -3-hydroxy-4-methyl-6- (3-methoxycarbonyl) benzyl-N-Menle was added]-1-Cyclosporin (1.60g, 1.20mmol) in tetrahydrofuran (5 ml). The mixture was stirred at-78 ℃ for three hours. After bubbling carbon dioxide gas into the reaction mixture for 15 minutes, the mixture was stirred at-78 ℃ for another 1 hour. The cooling bath was then removed and the reaction mixture was allowed to warm to room temperature to allow unreacted carbon dioxide to vent. The mixture was cooled to-78 ℃ and chloromethyl chloroformate (2.00ml) was added. The mixture was stirred and allowed to warm to room temperature overnight. Brine (5ml) was added to quench the reaction. Most of the tetrahydrofuran was removed under reduced pressure. Ethyl acetate (50ml) and brine (50ml) were added and the mixture was separated. The organic layer was dried over magnesium sulfate and evaporated under reduced pressure. The residue was purified by chromatography (hexane/acetone) to give 0.60g of pure [ (3R,4R) -3-hydroxy-4-methyl-6- (3- (N, N-diisopropylamino) carbonyl) benzyl-N-Menle]-1- [ α -methylene-Sar]-3-cyclosporin [ formula: c₇₅H₁₂₈N₁₂O₁₃(ii) a Accurate quality: 1404.97, respectively; MS (M/z) 1405.67(M +1)⁺]。

[ (3R,4R) -3-hydroxy-4-methyl-6- (3- (N, N-diisopropylamino) carbonyl) benzyl-N-Menle ] -1- [ (S) - (4-hydroxybutylthio) methyl-Sar ] -3-cyclosporin

To [ (3R,4R) -3-hydroxy-4-methyl-6- (3- (N, N-diisopropylamino) carbonyl) benzyl-N-Menle]-1- [ α -methylene-Sar]To a solution of (0.60g, 0.43mmol) of (E) -3-cyclosporin in methanol (25ml) were added (0.45g, 4.25mmol) of 4-mercapto-1-butanol and (0.10g, 4.25mmol) of lithium hydroxide. The reaction mixture was stirred at room temperature overnight. ReducingMost of the methanol was evaporated under pressure. Ethyl acetate (60ml) and brine (30ml) were added and the mixture was separated. The organic layer was dried over magnesium sulfate and evaporated under reduced pressure. The residue was purified by chromatography (dichloromethane/methanol) to give pure [ (3R,4R) -3-hydroxy-4-methyl-6- (3- (N, N-diisopropylamino) carbonyl) benzyl-N-Menle]-1- [ (S) - (4-hydroxybutylthio) methyl-Sar]-3-cyclosporin [ formula: c₇₉H₁₃₈N₁₂O₁₄S; accurate quality: 1511.02, respectively; MS (M/z) 1511.70(M +1)⁺(ii) a HPLC RT: 17.00 min. (C8 reverse phase column: 250 mm; acetonitrile/water (0.05% trifluoroacetic acid); operating temperature: 64 ℃ C.; detector: 210nm)]。

Example 6

[ (3R,4R) -3-hydroxy-4-methyl-6- (3-carboxy) benzyl-N-Menle]-1- [ (S) - (4-hydroxybutylthio) methyl-Sar]-3-Cyclosporin

[ (3R,4R) -3-hydroxy-4-methyl-6- (3-carboxy) benzyl-N-Menle ] -1- [ α -methylene-Sar ] -3-cyclosporin

N-butyllithium (2.65M, 7.7ml, 20.41mmol) was added to a solution of diisopropylamine (2.05g, 20.30mmol) in tetrahydrofuran (50ml) at-78 ℃ under nitrogen. After stirring the reaction mixture for 1 hour, [ (3R,4R) -3-hydroxy-4-methyl-6- (3-carboxy) benzyl-N-Menle was added]-1-Cyclosporin (2.40g, 1.20mmol) in tetrahydrofuran (10 ml). The mixture was stirred at-78 ℃ for three hours. After bubbling carbon dioxide gas into the reaction mixture for 15 minutes, the mixture was stirred at-78 ℃ for another 1 hour. The cooling bath was then removed and the reaction mixture was allowed to warm to room temperature to allow unreacted carbon dioxide to vent. The mixture was cooled to-78 ℃ and chloromethyl chloroformate (3.00ml) was added. The mixture was stirred and allowed to warm to room temperature overnight. Brine (5ml) was added to quench the reaction. Most of the tetrahydrofuran was removed under reduced pressure. Ethyl acetate (8) was added0ml) and brine (50ml), and the mixture was separated. The organic layer was dried over magnesium sulfate and evaporated under reduced pressure to give 1.50g of a crude product [ (3R,4R) -3-hydroxy-4-methyl-6- (3-carboxy) benzyl-N-Menle]-1- [ α -methylene-Sar]-3-cyclosporin [ formula: c₆₉H₁₁₅N₁₁O₁₄(ii) a Accurate quality: 1321.86, respectively; MS (M/z) 1322.45(M +1)⁺]。

[ (3R,4R) -3-hydroxy-4-methyl-6- (3-methoxycarbonyl) benzyl-N-Menle ] -1- [ α -methylene-Sar ] -3-cyclosporin

To [ (3R,4R) -3-hydroxy-4-methyl-6- (3-carboxy) benzyl-N-Menle]-1- [ α -methylene-Sar]To a solution of (1.32g, 1.00mmol) of (E) -3-cyclosporin in N, N-dimethylformamide (15ml) were added potassium carbonate (0.38g, 2.75mmol) and methyl iodide (0.50g, 3.52 mmol). The mixture was stirred at room temperature for three hours. Ethyl acetate (100ml) and water (50ml) were added to the solution, and the mixture was separated. The organic layer was dried over magnesium sulfate and evaporated under reduced pressure. The residue was purified by chromatography (dichloromethane/methanol) to give 1.60g of pure [ (3R,4R) -3-hydroxy-4-methyl-6- (3-methoxycarbonyl) benzyl-N-Menle]-1- [ α -methylene-Sar]-3-cyclosporin [ formula: c₇₀H₁₁₇N₁₁O₁₄(ii) a Accurate quality: 1335.88, respectively; MS (M/z) 1336.64(M +1)⁺]。

[ (3R,4R) -3-hydroxy-4-methyl-6- (3-methoxycarbonyl) benzyl-N-Menle ] -1- [ (S) - (4-hydroxybutylthio) methyl-Sar ] -3-cyclosporin

To [ (3R,4R) -3-hydroxy-4-methyl-6- (3-methoxycarbonyl) benzyl-N-Menle]-1- [ α -methylene-Sar]To a solution of (0.60g, 0.45mmol) of (E) -3-cyclosporin in methanol (25ml) were added (0.45g, 4.25mmol) of 4-mercapto-1-butanol and (0.10g, 4.25mmol) of lithium hydroxide.The reaction mixture was stirred at room temperature overnight. Most of the methanol was evaporated under reduced pressure. Ethyl acetate (60ml) and brine (30ml) were added and the mixture was separated. The organic layer was dried over magnesium sulfate and evaporated under reduced pressure. The residue was purified by chromatography (dichloromethane/methanol) to give pure [ (3R,4R) -3-hydroxy-4-methyl-6- (3-methoxycarbonyl) benzyl-N-Menle]-1- [ (S) - (4-hydroxybutylthio) methyl-Sar]-3-cyclosporin [ formula: c₇₄H₁₂₇N₁₁O₁₅S; accurate quality: 1441.92, respectively; MS (M/z) 1422.65(M +1)⁺]。

Example 7

[ (3R,4R) -3-hydroxy-4-methyl-6- (3-carboxy) benzyl-N-Menle]-1- [ (S) - (4-hydroxybutylthio) methyl-Sar]-3-Cyclosporin

To [ (3R,4R) -3-hydroxy-4-methyl-6- (3-methoxycarbonyl) benzyl-N-Menle]-1- [ (S) - (4-hydroxybutylthio) methyl-Sar]To a solution of-3-cyclosporin (85mg, 0.06mmol) in methanol (5ml) was added a solution of lithium hydroxide (15mg) in water (5 ml). The mixture was stirred at room temperature overnight. Most of the methanol was evaporated under reduced pressure. The pH of the aqueous mixture was adjusted to 6 with 1.0N hydrochloric acid. Ethyl acetate (60ml) and brine (10ml) were added to the solution, and the mixture was separated. The organic layer was dried over magnesium sulfate and evaporated under reduced pressure. Purifying the residue to obtain pure [ (3R,4R) -3-hydroxy-4-methyl-6- (3-carboxy) benzyl-N-Menle]-1- [ (S) - (4-hydroxybutylthio) methyl-Sar]-3-cyclosporin [ formula: c₇₃H₁₂₅N₁₁O₁₅S; accurate quality: 1427.91, respectively; MS (M/z) 1428.63(M +1)⁺(ii) a HPLC RT: 14.37 min. (C8 reverse phase column: 250 mm; acetonitrile/water (0.05% trifluoroacetic acid); operating temperature: 64 ℃ C.; detector: 210nm)]。

Example 8

[ (3R,4R) -3-hydroxy-4-methyl-6- (2-hydroxyethyl) -N-Menle]-1- [ (S) - (4-hydroxybutylthio) methyl base-Sar]-3-Cyclosporin

[ (3R,4R) -3-acetoxy-4-methyl- (6-methoxycarbonylmethylene) -N-Menle ] -1-Cyclosporin

To a solution of methoxycarbonylmethylenetriphenylphosphorane (6.20g, 18.54mmol) in anhydrous tetrahydrofuran (100ml) under nitrogen was added [ (3R,4R) -3-acetoxy-4-methyl-6-oxo-N-Menle]-1-Cyclosporin (4.10g, 3.33 mmol). The mixture was stirred and heated to reflux for six hours. Saturated ammonium chloride solution (20ml) was then added to quench the reaction. Most of the tetrahydrofuran was evaporated under reduced pressure. Ethyl acetate (150ml) and brine (50ml) were added and the mixture was separated. The organic layer was dried over magnesium sulfate and evaporated under reduced pressure. The residue was purified by chromatography (dichloromethane/methanol) to give 3.60g of pure [ (3R,4R) -3-acetoxy-4-methyl- (6-methoxycarbonylmethylene) -N-Menle]-1-cyclosporin [ formula: c₆₅H₁₁₃N₁₁O₁₅(ii) a Accurate quality: 1287.84, respectively; MS (M/z) 1288.61(M +1)⁺(ii) a HPLC RT: 16.43 min. (C8 reverse phase column: 250 mm; acetonitrile/water (0.05% trifluoroacetic acid); operating temperature: 64 ℃ C.; detector: 210nm)]。

[ (3R,4R) -3-acetoxy-4-methyl-6- (methoxycarbonylmethyl) -N-Menle ] -1-Cyclosporin

To [ (3R,4R) -3-acetoxy-4-methyl-6- (methoxycarbonylmethylene) -N-Menle]To a solution of-1-cyclosporin (3.80g, 2.95mmol) in methanol (100ml) was added palladium (10 wt%, on carbon, 250mg) and acetic acid (5 drops). The mixture was stirred at room temperature under hydrogen for eight hours. Filtering and mixingThe filtrate was evaporated under reduced pressure to give crude [ (3R,4R) -3-acetoxy-4-methyl-6- (methoxycarbonylmethyl) -N-Menle]-1-cyclosporin [ formula: c₆₅H₁₁₅N₁₁O₁₅(ii) a Accurate quality: 1289.86, respectively; MS (M/z) 1290.59(M +1)⁺(ii) a HPLC RT: 16.50 min. (C8 reverse phase column: 250 mm; acetonitrile/water (0.05% trifluoroacetic acid); operating temperature: 64 ℃ C.; detector: 210nm)]。

[ (3R,4R) -3-hydroxy-4-methyl-6- (2-hydroxyethyl) -N-Menle ] -1-Cyclosporin

To [ (3R,4R) -3-acetoxy-4-methyl-6- (methoxycarbonylmethyl) -N-Menle]To a solution of (3.50g, 2.71mmol) of (E) -1-cyclosporin in methanol (100ml) was added cesium chloride (5.00g, 29.78 mmol). Sodium borohydride (10.00g, 262.95mmol) was then added portionwise over 30 minutes. After stirring the mixture at room temperature for a further eight hours, most of the methanol was evaporated under reduced pressure. Ethyl acetate (150ml) and saturated sodium bicarbonate solution (150ml) were added and the mixture was separated. The organic layer was washed with brine, dried over magnesium sulfate and evaporated under reduced pressure. The residue was purified by chromatography (dichloromethane/methanol) to give 2.85g of [ (3R,4R) -3-hydroxy-4-methyl-6- (2-hydroxyethyl) -N-Menle]-1-cyclosporin [ formula: c₆₂H₁₁₃N₁₁O₁₃(ii) a Accurate quality: 1219.85, respectively; MS (M/z) 1220.68(M +1)⁺(ii) a HPLC RT: 15.80 min. (C8 reverse phase column: 250 mm; acetonitrile/water (0.05% trifluoroacetic acid); operating temperature: 64 ℃ C.; detector: 210nm)]。

[ (3R,4R) -3-hydroxy-4-methyl-6- (2-hydroxyethyl) -N-Menle ] -1- [ α -methylene-Sar ] -3-cyclosporin

N-butyllithium (2.65M, 9.60ml, 25.30 mmo) was added at-78 ℃ under nitrogenl) was added to a solution of diisopropylamine (2.56g, 25.30mmol) in tetrahydrofuran (80 ml). After stirring the reaction mixture for 1 hour, [ (3R,4R) -3-hydroxy-4-methyl-6- (2-hydroxyethyl) -N-Menle was added]-1-Cyclosporin (2.85g, 2.30mmol) in tetrahydrofuran (15 ml). The mixture was stirred at-78 ℃ for three hours. After bubbling carbon dioxide gas into the reaction mixture for 15 minutes, the mixture was stirred at-78 ℃ for another 1 hour. The cooling bath was then removed and the reaction mixture was allowed to warm to room temperature to allow unreacted carbon dioxide to vent. The mixture was cooled to-78 ℃ and chloromethyl chloroformate (3.00ml) was added. The mixture was stirred and allowed to warm to room temperature overnight. Brine (5ml) was added to quench the reaction. Most of the tetrahydrofuran was removed under reduced pressure. Ethyl acetate (80ml) and brine (50ml) were added and the mixture was separated. The organic layer was dried over magnesium sulfate and evaporated under reduced pressure. The residue was purified by chromatography (dichloromethane/methanol) to give 1.0g of the product [ (3R,4R) -3-hydroxy-4-methyl-6- (2-hydroxyethyl) -N-Menle]-1- [ α -methylene-Sar]-3-cyclosporin [ formula: c₆₃H₁₁₃N₁₁O₁₃(ii) a Accurate quality: 1231.85, respectively; MS (M/z) 1232.55(M +1)⁺(ii) a HPLC RT: 15.14 min. (C8 reverse phase column: 250 mm; acetonitrile/water (0.05% trifluoroacetic acid); operating temperature: 64 ℃ C.; detector: 210nm)]。

[ (3R,4R) -3-hydroxy-4-methyl-6- (2-hydroxyethyl) -N-Menle ] -1- [ (S) - (4-hydroxybutylthio) methyl-Sar ] -3-cyclosporin

To [ (3R,4R) -3-hydroxy-4-methyl-6- (2-hydroxyethyl) -N-Menle]-1- [ α -methylene-Sar]To a solution of (0.60g, 0.45mmol) of (E) -3-cyclosporin in methanol (50ml) were added (0.45g, 4.25mmol) of 4-mercapto-1-butanol and (0.10g, 4.25mmol) of lithium hydroxide. The reaction mixture was stirred at room temperature overnight. Most of the methanol was evaporated under reduced pressure. Ethyl acetate (60ml) and brine (30ml) were added and the mixture was separated. The organic layer was dried over magnesium sulfate and evaporated under reduced pressure. The residue was purified by chromatography (dichloromethane/methanol) to give pure product[ (3R,4R) -3-hydroxy-4-methyl-6- (2-hydroxyethyl) -N-Menle]-1- [ (S) - (4-hydroxybutylthio) methyl-Sar]-3-cyclosporin [ formula: c₆₇H₁₂₃N₁₁O₁₄S; accurate quality: 1337.90, respectively; MS (M/z) 1338.74(M +1)⁺(ii) a HPLC RT: 12.98 min. (C8 reverse phase column: 250 mm; acetonitrile/water (0.05% trifluoroacetic acid); operating temperature: 64 ℃ C.; detector: 210nm)]。

Example 9

[8- (2-Acetylaminoethyl) -6, 7-dihydro-MeBmt ] -1- [ (R) -2-methoxymethyl-Sar ] -3-Cyclosporin

(3-cyanopropyl) triphenyl phosphine bromide

4-Bromobutyronitrile (10ml, 0.10mol) and triphenylphosphine (36.23g, 0.10mol) were added to toluene (200 ml). The mixture was stirred and heated to reflux overnight. After cooling to room temperature, the precipitate was filtered off, washed with toluene and hexane and dried in vacuo to give 26.60g of product. [ molecular formula: c₂₂H₂₁NP⁺(ii) a Accurate quality: 330.14, respectively; MS (M/z) 330.23(M)⁺]。

[ 8-cyanomethyl-3-acetyl-MeBmt ] -1-cyclosporin

To a dry flask were added (3-cyanopropyl) triphenyl phosphonium bromide (7.98g, 19.50mmol) and anhydrous tetrahydrofuran (60ml) under nitrogen. The reaction mixture was placed in an ice-water bath and sodium tert-butoxide (2.19g, 22.75mmol) was added. After stirring the mixture for 2 hours [ (3R,4R) -3-acetoxy-4-methyl-6-oxo-N-Menle was added]-a solution of 1-cyclosporin (4.00g, 3.25mmol) in anhydrous tetrahydrofuran (20 ml). Will be mixed withThe mixture was stirred at 0 ℃ for a further 5 hours. Saturated ammonium chloride solution (20ml) was then added to quench the reaction. Most of the tetrahydrofuran was evaporated under reduced pressure. Ethyl acetate (100ml) and brine (100ml) were added and the mixture was separated. The organic layer was dried over magnesium sulfate and evaporated under reduced pressure. The residue was purified by chromatography (dichloromethane/methanol) to give 2.00g of pure [ 8-cyanomethyl-3-acetyl-MeBmt]-1-cyclosporin [ formula: c₆₆H₁₁₄N₁₂O₁₃(ii) a Accurate quality: 1282.86, respectively; MS (M/z) 1283.51(M +1)⁺,1305.73(M+Na)⁺(ii) a HPLC RT: 16.50 min. (C8 reverse phase column: 250 mm; acetonitrile/water (0.05% trifluoroacetic acid); operating temperature: 64 ℃ C.; detector: 210nm)]。

[ 8-cyanomethyl-MeBmt ] -1-cyclosporin

Reacting [ 8-cyanomethyl-3-acetyl-MeBmt]-1-Cyclosporin (2.00g, 1.56mmol) was dissolved in methanol (20 ml). Water (10ml) and tetramethylammonium hydroxide pentahydrate (0.85g, 4.68mmol) were added. The mixture was stirred at room temperature for 2 hours. Most of the methanol is then evaporated. Ethyl acetate (100ml) and brine (100ml) were added and the mixture was separated. The organic layer was dried over magnesium sulfate and evaporated under reduced pressure. The residue was purified by chromatography (dichloromethane/methanol) to give 1.70g of pure [ 8-cyanomethyl-MeBmt]-1-cyclosporin [ formula: c₆₄H₁₁₂N₁₂O₁₂(ii) a Accurate quality: 1240.85, respectively; MS (M/z) 1241.54(M +1)⁺,1263.73(M+Na)⁺(ii) a HPLC RT: 14.80 min. (C8 reverse phase column: 250 mm; acetonitrile/water (0.05% trifluoroacetic acid); operating temperature: 64 ℃ C.; detector: 210nm)]。

[8- (2-aminoethyl) -6, 7-dihydro-MeBmt ] -1-Cyclosporin

To [ 8-cyanomethyl-MeBmt under nitrogen]To a solution of (2.00g, 1.61mmol) of (E) -1-cyclosporin in methanol (50ml) was added nickel (II) chloride hexahydrate (0.19g, 0.81 mmol). The reaction mixture was placed in an ice-water bath. Sodium borohydride (3.05g, 80.50mmol) was added in four portions over a two hour period. After the mixture was stirred at 0 ℃ for another 2 hours, water (10ml) was added. Most of the methanol was evaporated under reduced pressure. Ethyl acetate (50ml) and saturated sodium bicarbonate solution (50ml) were added to the solution, and the mixture was separated. The organic layer was washed with brine, dried over magnesium sulfate and evaporated under reduced pressure. The residue was dissolved in methanol (30 ml). Palladium (10 wt% on carbon, 20mg) and acetic acid (5 drops) were added. The mixture was stirred at room temperature under hydrogen for 2 hours. The mixture was filtered and the filtrate was evaporated under reduced pressure to give crude [8- (2-aminoethyl) -6, 7-dihydro-MeBmt]-1-cyclosporin [ formula: c₆₄H₁₁₈N₁₂O₁₂(ii) a Accurate quality: 1246.90, respectively; MS (M/z) 1247.69(M +1)⁺(ii) a HPLC RT: 11.13 min. (C8 reverse phase column: 250 mm; acetonitrile/water (0.05% trifluoroacetic acid); operating temperature: 64 ℃ C.; detector: 210nm)]。

[8- (2- (tert-Butoxycarbonyl) aminoethyl) -6, 7-dihydro-MeBmt ] -1-Cyclosporin

Reacting [8- (2-aminoethyl) -6, 7-dihydro-MeBmt]-1-Cyclosporin (crude, 1.61mmol) was dissolved in tetrahydrofuran (20 ml). Saturated sodium bicarbonate solution (20ml) and di-tert-butyl dicarbonate (0.39g, 1.77mmol) were added. The mixture was stirred at room temperature for 2 hours. Most of the tetrahydrofuran was evaporated under reduced pressure. Ethyl acetate (30ml) and brine (30ml) were added and the mixture was separated. The organic layer was dried over magnesium sulfate and evaporated under reduced pressure. The residue was purified by chromatography (hexane/acetone) to give 2.00g of pure [8- (2- (tert-butoxycarbonyl) aminoethyl) -6, 7-dihydro-MeBmt]-1-cyclosporin [ formula: c₆₉H₁₂₆N₁₂O₁₄(ii) a Accurate quality: 1346.95, respectively; MS (M/z) 1347.54(M +1)⁺,1369.78(M+Na)⁺；HPLC RT：17.08 min. (C8 reverse phase column: 250 mm; acetonitrile/water (0.05% trifluoroacetic acid); operating temperature: 64 ℃ C.; detector: 210nm)]。

[8- (2- (tert-Butoxycarbonyl) aminoethyl) -6, 7-dihydro-MeBmt ] -1- [ (S) -2-methoxycarbonyl-Sar ] -3-Cyclosporin

N-butyllithium (2.20M, 3.38ml, 7.42mmol) was added to a solution of diisopropylamine (1.06ml, 7.42mmol) in tetrahydrofuran (20ml) at-78 ℃ under nitrogen. After stirring the reaction mixture for 1 hour, [8- (2- (tert-butoxycarbonyl) aminoethyl) -6, 7-dihydro-MeBmt was added over 10 minutes]-1-Cyclosporin (1.00g, 0.74mmol) in tetrahydrofuran (10 ml). The mixture was stirred at-78 ℃ for 2 hours. After bubbling carbon dioxide gas into the reaction mixture for 20 minutes, the mixture was stirred at-78 ℃ for another 1 hour. The cooling bath was then removed and the reaction mixture was allowed to slowly warm to 0 ℃. Most of the tetrahydrofuran was removed at room temperature under vacuum. The residue was quenched by addition of saturated citric acid solution, the pH of the aqueous layer was adjusted to 3-4 with 1N hydrochloric acid, and the precipitated oil was extracted with ethyl acetate (100 ml). The aqueous layer was extracted with ethyl acetate (100 ml. times.3). The combined ethyl acetate layers were washed with brine, dried over magnesium sulfate and evaporated under reduced pressure to give 0.50g of [8- (2- (tert-butoxycarbonyl) aminoethyl) -6, 7-dihydro-MeBmt]-1- [ (S) -2-carboxy-Sar]-3-cyclosporin [ formula: c₇₀H₁₂₆N₁₂O₁₆(ii) a Accurate quality: 1390.94, respectively; MS (M/z) 1391.60(M +1)⁺(ii) a HPLC RT: 15.57 min. (C8 reverse phase column: 250 mm; acetonitrile/water (0.05% trifluoroacetic acid); operating temperature: 64 ℃ C.; detector: 210nm)]。

Reacting [8- (2- (tert-butoxycarbonyl) aminoethyl) -6, 7-dihydro-MeBmt]-1- [ (S) -2-carboxy-Sar]-3-Cyclosporin (0.50g, 0.36mmol) was dissolved in acetone (10 ml). Methyl iodide (0.03ml, 0.54mmol) and potassium carbonate (0.08g, 0.54mmol) were added. The mixture was stirred at room temperature for 2 hours. Most of the acetone was evaporated under reduced pressure. Then theEthyl acetate (40ml) and water (40ml) were added to the solution, and the mixture was separated. The ethyl acetate layer was dried over magnesium sulfate and evaporated under reduced pressure to give 0.51g of crude [8- (2- (tert-butoxycarbonyl) aminoethyl) -6, 7-dihydro-MeBmt]-1- [ (S) -2-methoxycarbonyl-Sar]-3-cyclosporin [ formula: c₇₁H₁₂₈N₁₂O₁₆(ii) a Accurate quality: 1404.96, respectively; MS (M/z) 1405.64(M +1)⁺,1427.86(M+Na)⁺(ii) a HPLC RT: 16.65 min. (C8 reverse phase column: 250 mm; acetonitrile/water (0.05% trifluoroacetic acid); operating temperature: 64 ℃ C.; detector: 210nm)]。

[8- (2- (tert-Butoxycarbonyl) aminoethyl) -6, 7-dihydro-MeBmt ] -1- [ (R) -2-hydroxymethyl-Sar ] -3-Cyclosporin

Reacting [8- (2- (tert-butoxycarbonyl) aminoethyl) -6, 7-dihydro-MeBmt]-1- [ (S) -2-methoxycarbonyl-Sar]-3-Cyclosporin (0.51g, 0.36mmol) was dissolved in tetrahydrofuran (10 ml). Sodium borohydride (0.68g, 18.00mmol) and cesium chloride (0.49g, 1.51mmol) were added. The mixture was stirred at room temperature, and methanol (10ml) was added dropwise over 1 hour. The mixture was stirred for an additional 2 hours. Most of the solvent was evaporated under reduced pressure. Ethyl acetate (40ml) and water (40ml) were then added and the mixture was separated. The ethyl acetate layer was dried over magnesium sulfate and evaporated under reduced pressure to give 0.50g of crude [8- (2- (tert-butoxycarbonyl) aminoethyl) -6, 7-dihydro-MeBmt]-1- [ (R) -2-hydroxymethyl-Sar]-3-cyclosporin [ formula: c₇₀H₁₂₈N₁₂O₁₅(ii) a Accurate quality: 1376.96, respectively; MS (M/z) 1377.71(M +1)⁺,1388.85(M+Na)⁺(ii) a HPLC RT: 16.49 min. (C8 reverse phase column: 250 mm; acetonitrile/water (0.05% trifluoroacetic acid); operating temperature: 64 ℃ C.; detector: 210nm)]。

[8- (2-Acetylaminoethyl) -6, 7-dihydro-MeBmt ] -1- [ (R) -2-hydroxymethyl-Sar ] -3-Cyclosporin

Reacting [8- (2- (tert-butoxycarbonyl) aminoethyl) -6, 7-dihydro-MeBmt]-1- [ (R) -2-hydroxymethyl-Sar]-3-Cyclosporin (0.50g, 0.36mmol) was dissolved in dichloromethane (10ml) and placed in an ice-water bath. Trifluoroacetic acid (5ml) was added. The mixture was stirred at room temperature for one hour. Another portion of dichloromethane (20ml) was added. The mixture was washed with brine (30ml), saturated sodium bicarbonate solution (30ml), dried over magnesium sulfate and evaporated under reduced pressure to give crude [8- (2-aminoethyl) -6, 7-dihydro-MeBmt]-1- [ (R) -2-hydroxymethyl-Sar]-3-cyclosporin [ formula: c₆₅H₁₂₀N₁₂O₁₃(ii) a Accurate quality: 1276.91, respectively; MS (M/z) 1277.75(M +1)⁺(ii) a HPLC RT: 9.13 min. (C8 reverse phase column: 250 mm; acetonitrile/water (0.05% trifluoroacetic acid); operating temperature: 64 ℃ C.; detector: 210nm)]. Crude [8- (2-aminoethyl) -6, 7-dihydro-MeBmt]-1- [ (R) -2-hydroxymethyl-Sar]-3-Cyclosporin was dissolved in dichloromethane (10 ml). Acetic acid (0.12ml, 2.20mmol), HBTU (0.41g, 1.08mmol), 1-hydroxybenzotriazole (0.15g, 1.08mmol) and pyridine (0.50ml) were added. The mixture was stirred at room temperature for 2 hours. Dichloromethane (30ml) and brine (50ml) were then added and separated. The dichloromethane layer was dried over magnesium sulfate and evaporated under reduced pressure. The residue was purified by chromatography (dichloromethane/methanol) to give 27mg of pure [8- (2-acetamidomethyl) -6, 7-dihydro-MeBmt]-1- [ (R) -2-hydroxymethyl-Sar]-3-cyclosporin [ formula: c₆₇H₁₂₂N₁₂O₁₄(ii) a Accurate quality: 1318.92, respectively; MS (M/z) 1319.72(M +1)⁺,1341.91(M+Na)⁺(ii) a HPLC RT: 14.12 min. (C8 reverse phase column: 250 mm; acetonitrile/water (0.05% trifluoroacetic acid); operating temperature: 64 ℃ C.; detector: 210nm)]。

[8- (2-Acetylaminoethyl) -6, 7-dihydro-MeBmt ] -1- [ (R) -2-methoxymethyl-Sar ] -3-Cyclosporin

Reacting [8- (2-acetamido)Ylethyl) -6, 7-dihydro-MeBmt]-1- [ (R) -2-hydroxymethyl-Sar]-3-Cyclosporin (0.20g, 0.15mmol) was dissolved in benzene (10 ml). Methyl iodide (0.21g, 1.52mmol), tetra-n-butylammonium bromide (0.49g, 1.52mmol), sodium hydroxide (1.00g, 2.50mmol) and water (2ml) were added. The mixture was stirred at room temperature overnight. Ethyl acetate (50ml) and brine (50ml) were then added and separated. The ethyl acetate layer was dried over magnesium sulfate and evaporated under reduced pressure. The residue was purified by chromatography (dichloromethane/methanol) to give 60mg of pure [8- (2-acetamidomethyl) -6, 7-dihydro-MeBmt]-1- [ (R) -2-methoxymethyl-Sar]-3-cyclosporin [ formula: c₆₈H₁₂₄N₁₂O₁₄(ii) a Accurate quality: 1332.94, respectively; MS (M/z) 1333.69(M +1)⁺,1355.86(M+Na)⁺(ii) a HPLC RT: 16.18 min. (C8 reverse phase column: 250 mm; acetonitrile/water (0.05% trifluoroacetic acid); operating temperature: 64 ℃ C.; detector: 210nm)]。

Example 10

[8- (2-Acetylaminoethyl) -6, 7-dihydro-MeBmt]-1- [ (S) - (4-hydroxybutylthio) methyl-Sar]-3- Cyclosporin

[8- (2- (tert-Butoxycarbonyl) aminoethyl) -6, 7-dihydro-MeBmt ] -1- [ α -methylene-Sar ] -3-Cyclosporin

N-butyllithium (2.20M, 6.75ml, 14.85mmol) was added to a solution of diisopropylamine (2.11ml, 14.85mmol) in tetrahydrofuran (40ml) at-78 ℃ under nitrogen. After stirring the reaction mixture for 1 hour, [8- (2- (tert-butoxycarbonyl) aminoethyl) -6, 7-dihydro-MeBmt was added over 10 minutes]-1-Cyclosporin (2.00g, 1.48mmol) in tetrahydrofuran (15 ml). The mixture was stirred at-78 ℃ for 2 hours. After bubbling carbon dioxide gas into the reaction mixture for 30 minutes, the mixture was stirred at-78 ℃ for another 1 hour. The cooling bath was then removed and the reaction mixture was allowed to warm to room temperature to allow for non-reversionThe corresponding carbon dioxide is discharged. The mixture was cooled to-78 ℃ and chloromethyl chloroformate (1.32ml, 14.85mmol) was added. The mixture was stirred and allowed to warm to room temperature overnight. Brine (5ml) was added to quench the reaction. Most of the tetrahydrofuran was removed under reduced pressure. Ethyl acetate (50ml) and brine (50ml) were added and the mixture was separated. The organic layer was dried over magnesium sulfate and evaporated under reduced pressure. The residue was purified by chromatography (hexane/acetone) to give 0.60g of pure [8- (2- (tert-butoxycarbonyl) aminoethyl) -6, 7-dihydro-MeBmt]-1- [ α -methylene-Sar]-3-cyclosporin [ formula: c₇₀H₁₂₆N₁₂O₁₄(ii) a Accurate quality: 1358.95, respectively; MS (M/z) 1359.59(M +1)⁺,1381.81(M+Na)⁺(ii) a HPLC RT: 18.44 min. (C8 reverse phase column: 250 mm; acetonitrile/water (0.05% trifluoroacetic acid); operating temperature: 64 ℃ C.; detector: 210nm)]。

[8- (2-Acetylaminoethyl) -6, 7-dihydro-MeBmt ] -1- [ α -methylene-Sar ] -3-Cyclosporin

Reacting [8- (2- (tert-butoxycarbonyl) aminoethyl) -6, 7-dihydro-MeBmt]-1- [ α -methylene-Sar]-3-Cyclosporin (0.60g, 0.44mmol) was dissolved in dichloromethane (10ml) and placed in an ice-water bath. Trifluoroacetic acid (5ml) was added. The mixture was stirred at room temperature for one hour. Another portion of dichloromethane (20ml) was added. The mixture was washed with brine (30ml), saturated sodium bicarbonate solution (30ml), dried over magnesium sulfate and evaporated under reduced pressure to give crude [8- (2-aminoethyl) -6, 7-dihydro-MeBmt]-1- [ α -methylene-Sar]-3-cyclosporin [ formula: c₆₅H₁₁₈N₁₂O₁₂(ii) a Accurate quality: 1258.90, respectively; MS (M/z) 1259.77(M +1)⁺,1281.84(M+Na)⁺(ii) a HPLC RT: 13.00 min. (C8 reverse phase column: 250 mm; acetonitrile/water (0.05% trifluoroacetic acid); operating temperature: 64 ℃ C.; detector: 210nm)]. Crude [8- (2-aminoethyl) -6, 7-dihydro-MeBmt]-1- [ α -methylene-Sar]-3-Cyclosporin was dissolved in dichloromethane (10 ml). Acetic acid (0.12ml, 2.20mmol), HBTU (0.50g, 1.32mmol) were addedmmol), 1-hydroxybenzotriazole (0.18g, 1.32mmol) and pyridine (1 ml). The mixture was stirred at room temperature for one hour. Dichloromethane (30ml) and brine (50ml) were then added and separated. The dichloromethane layer was dried over magnesium sulfate and evaporated under reduced pressure. The residue was purified by chromatography (dichloromethane/methanol) to give 0.30g of pure [8- (2-acetamidomethyl) -6, 7-dihydro-MeBmt]-1- [ α -methylene-Sar]-3-cyclosporin [ formula: c₆₅H₁₁₈N₁₂O₁₂(ii) a Accurate quality: 1258.90, respectively; MS (M/z) 1259.77(M +1)⁺,1281.84(M+Na)⁺(ii) a HPLC RT: 13.00 min. (C8 reverse phase column: 250 mm; acetonitrile/water (0.05% trifluoroacetic acid); operating temperature: 64 ℃ C.; detector: 210nm)]。

[8- (2-Acetylaminoethyl) -6, 7-dihydro-MeBmt ] -1- [ (S) - (4-Hydroxybutylsulfanyl) methyl-Sar ] -3-Cyclosporin

To [8- (2-acetamidoethyl) -6, 7-dihydro-MeBmt]-1- [ α -methylene-Sar]To a solution of (0.30g, 0.23mmol) of (E) -3-cyclosporin in methanol (10ml) were added (0.14ml, 1.38mmol) of 4-mercapto-1-butanol and (0.06g, 2.31mmol) of lithium hydroxide. The reaction mixture was stirred at room temperature for 5 hours. Most of the methanol was evaporated under reduced pressure. Ethyl acetate (30ml) and brine (30ml) were added and the mixture was separated. The organic layer was dried over magnesium sulfate and evaporated under reduced pressure. The residue was purified by chromatography (hexane/acetone from 90/10 to 75/25) to yield 14mg of pure [8- (2-acetamidomethyl) -6, 7-dihydro-MeBmt]-1- [ (S) - (4-hydroxybutylthio) methyl-Sar]-3-cyclosporin [ formula: c₇₁H₁₃₀N₁₂O₁₄S; accurate quality: 1406.95, respectively; MS (M/z) 1407.70(M +1)⁺,1429.89(M+Na)⁺(ii) a HPLC RT: 15.02 min. (C8 reverse phase column: 250 mm; acetonitrile/water (0.05% trifluoroacetic acid); operating temperature: 64 ℃ C.; detector: 210nm)]。

Example 11

[8- (2-hydroxyethyl) -6, 7-dihydro-MeBmt]-1- [ (S) - (4-hydroxybutylthio) methyl-Sar]-3-Cyclosporium sp Vegetable extract(4-ethoxy-4-oxobutyl) triphenylphosphine bromide

Ethyl 4-bromobutyrate (14.40ml, 0.10mol) and triphenylphosphine (26.29g, 0.10mol) were added to toluene (200 ml). The mixture was stirred and heated to reflux for 24 hours. After cooling to room temperature, the mixture was stirred at room temperature over the weekend. The precipitate was filtered off, washed with toluene and hexane and dried in vacuo to give 20.0g of product. [ molecular formula: c₂₄H₂₆O₂P⁺(ii) a Accurate quality: 377.17, respectively; MS (M/z) 377.20(M)⁺]。

[8- (2-ethoxy-2-oxoethyl) -3-acetyl-MeBmt ] -1-cyclosporin

To a dry flask were added (4-ethoxy-4-oxobutyl) triphenyl phosphonium bromide (8.88g, 19.48mmol) and anhydrous tetrahydrofuran (60ml) under nitrogen. The reaction mixture was placed in an ice-water bath and sodium tert-butoxide (2.18g, 22.74mmol) was added. After stirring the mixture for 2 hours [ (3R,4R) -3-acetoxy-4-methyl-6-oxo-N-Menle was added]-a solution of 1-cyclosporin (4.00g, 3.25mmol) in anhydrous tetrahydrofuran (20 ml). The mixture was stirred at 0 ℃ for a further 5 hours. Most of the tetrahydrofuran was evaporated under reduced pressure. Dichloromethane (100ml) and brine (100ml) were added and the mixture was separated. The organic layer was dried over magnesium sulfate and evaporated under reduced pressure. The residue was purified by chromatography (dichloromethane/methanol) to give 2.82g of [8- (2-ethoxy-2-oxoethyl) -3-acetyl-MeBmt]-1-cyclosporin [ formula: c₆₈H₁₁₉N₁₁O₁₅(ii) a Accurate quality: 1329.89, respectively; MS (M/z) 1330.51(M +1)⁺,1352.69(M+Na)⁺(ii) a HPLC RT: 17.72 min. (C8 reverse phase column: 250 mm; acetonitrile/water (0.05% trifluoroacetic acid); operating temperature: 64 ℃ C.; detector: 210nm)]。

[8- (2-hydroxyethyl) -MeBmt ] -1-Cyclosporin

Reacting [8- (2-ethoxy-2-oxoethyl) -3-acetyl-MeBmt]-1-Cyclosporin (2.82g, 2.12mmol) was dissolved in tetrahydrofuran (40 ml). Sodium borohydride (8.02g, 212.04mmol) and cesium chloride (3.56g, 21.20mmol) were added. The mixture was stirred at room temperature, and methanol (40ml) was added dropwise over 2 hours. The mixture was stirred for a further 3 hours. Most of the solvent was evaporated under reduced pressure. Ethyl acetate (100ml) and water (100ml) were then added and the mixture was separated. The ethyl acetate layer was dried over magnesium sulfate and evaporated under reduced pressure. The residue was purified by chromatography (dichloromethane/methanol) to give 2.70g of [8- (2-hydroxyethyl) -MeBmt]-1-cyclosporin [ formula: c₆₄H₁₁₅N₁₁O₁₃(ii) a Accurate quality: 1245.87, respectively; MS (M/z) 1246.56(M +1)⁺,1413.96(M+Na)⁺(ii) a HPLC RT: 14.15 min. (C8 reverse phase column: 250 mm; acetonitrile/water (0.05% trifluoroacetic acid); operating temperature: 64 ℃ C.; detector: 210nm)]。

[8- (2-hydroxyethyl) -6, 7-dihydro-MeBmt ] -1-cyclosporin

Reacting [8- (2-hydroxyethyl) -MeBmt]-1-Cyclosporin (2.70g, 2.17mmol) was dissolved in methanol (50 ml). Palladium (10 wt% on carbon, 50mg) and acetic acid (6 drops) were added. The mixture was stirred under hydrogen at room temperature overnight. The mixture was filtered and the filtrate was evaporated under reduced pressure to give 1.50g of crude [8- (2-hydroxyethyl) -6, 7-dihydro-MeBmt]-1-cyclosporin [ formula: c₆₄H₁₁₇N₁₁O₁₃(ii) a Extract of Chinese medicinal materialsThe quality is determined as follows: 1247.88, respectively; MS (M/z) 1248.66(M +1)⁺,1270.77(M+Na)⁺(ii) a HPLC RT: 14.32 min. (C8 reverse phase column: 250 mm; acetonitrile/water (0.05% trifluoroacetic acid); operating temperature: 64 ℃ C.; detector: 210nm)]。

[8- (2-hydroxyethyl) -6, 7-dihydro-MeBmt ] -1- [ α -methylene-Sar ] -3-Cyclosporin

N-butyllithium (2.65M,5.00ml, 13.23mmol) was added to a solution of diisopropylamine (1.87ml, 13.23mmol) in tetrahydrofuran (40ml) at-78 ℃ under nitrogen. After stirring the reaction mixture for 1 hour, add [8- (2-hydroxyethyl) -6, 7-dihydro-MeBmt in 10 minutes]-1-Cyclosporin (1.50g, 1.20mmol) in tetrahydrofuran (15 ml). The mixture was stirred at-78 ℃ for 2 hours. After bubbling carbon dioxide gas into the reaction mixture for 30 minutes, the mixture was stirred at-78 ℃ for another 1 hour. The cooling bath was then removed and the reaction mixture was allowed to warm to room temperature to allow unreacted carbon dioxide to vent. The mixture was cooled to-78 ℃ and chloromethyl chloroformate (1.07ml, 12.03mmol) was added. The mixture was stirred and allowed to warm to room temperature overnight. Brine (5ml) was added to quench the reaction. Most of the tetrahydrofuran was removed under reduced pressure. Ethyl acetate (50ml) and brine (50ml) were added and the mixture was separated. The organic layer was dried over magnesium sulfate and evaporated under reduced pressure. The residue was purified by chromatography (hexane/acetone) to give 0.50g of pure [8- (2-hydroxyethyl) -6, 7-dihydro-MeBmt]-1- [ α -methylene-Sar]-3-cyclosporin [ formula: c₆₅H₁₁₇N₁₁O₁₃(ii) a Accurate quality: 1259.88, respectively; MS (M/z) 1260.61(M +1)⁺,1282.78(M+Na)⁺(ii) a HPLC RT: 16.09 min. (C8 reverse phase column: 250 mm; acetonitrile/water (0.05% trifluoroacetic acid); operating temperature: 64 ℃ C.; detector: 210nm)]。

[8- (2-hydroxyethyl) -6, 7-dihydro-MeBmt ] -1- [ (S) - (4-hydroxybutylthio) methyl-Sar ] -3-cyclosporin

To [8- (2-hydroxyethyl) -6, 7-dihydro-MeBmt]-1- [ α -methylene-Sar]To a solution of (0.50g, 0.40mmol) of (E) -3-cyclosporin in methanol (10ml) were added (0.25ml, 2.38mmol) of 4-mercapto-1-butanol and (0.10g, 4.00mmol) of lithium hydroxide. The reaction mixture was stirred at room temperature for 4 hours. Most of the methanol was evaporated under reduced pressure. Ethyl acetate (30ml) and brine (30ml) were added and the mixture was separated. The organic layer was dried over magnesium sulfate and evaporated under reduced pressure. The residue was purified by chromatography (hexane/acetone) to give 80mg of pure [8- (2-hydroxyethyl) -6, 7-dihydro-MeBmt]-1- [ (S) - (4-hydroxybutylthio) methyl-Sar]-3-cyclosporin [ formula: c₆₉H₁₂₇N₁₁O₁₄S; accurate quality: 1365.93, respectively; MS (M/z) 1366.64(M +1)⁺,1388.84(M+Na)⁺(ii) a HPLC RT: 13.94 min. (C8 reverse phase column: 250 mm; acetonitrile/water (0.05% trifluoroacetic acid); operating temperature: 64 ℃ C.; detector: 210nm)]。

Example 12

[8- (2- (2-ethoxy-2-oxoethyl) amino) ethyl-6, 7-dihydro-MeBmt]-1- [ (S) - (4-Hydroxybutane) Thio) methyl-Sar]-3-cyclosporin (12a) and [8- (2-bis (2-ethoxy-2-oxoethyl) amino) ethyl-6, 7-bis Hydrogen-MeBmt]-1- [ (S) - (4-hydroxybutylthio) methyl-Sar]-3-Cyclosporin (12b)

[8- (2- (tert-Butoxycarbonyl) aminoethyl) -6, 7-dihydro-MeBmt ] -1- [ (S) - (4-hydroxybutylthio) methyl-Sar ] -3-cyclosporin

To [8- (2- (tert-butoxycarbonyl) aminoethyl) -6, 7-dihydro-MeBmt]-1- [ α -methylene-Sar]-3-Cyclosporin (0.25g, 0.18mmol) in methanol (15ml) 4-mercapto-1-butanol (0.12ml, 1.10mmol) and hydrogen were addedLithium oxide (0.04g, 1.84 mmol). The reaction mixture was stirred at room temperature overnight. Most of the methanol was evaporated under reduced pressure. Ethyl acetate (30ml) and brine (30ml) were added and the mixture was separated. The organic layer was dried over magnesium sulfate and evaporated under reduced pressure. The residue was purified by chromatography (hexane/acetone from 90/10 to 75/25) to yield 100mg of pure [8- (2- (tert-butoxycarbonyl) aminoethyl) -6, 7-dihydro-MeBmt]-1- [ (S) - (4-hydroxybutylthio) methyl-Sar]-3-cyclosporin [ formula: c₇₄H₁₃₆N₁₂O₁₅S; accurate quality: 1465.00, respectively; MS (M/z) 1465.53(M +1)⁺,1487.84(M+Na)⁺(ii) a HPLC RT: 16.77 min. (C8 reverse phase column: 250 mm; acetonitrile/water (0.05% trifluoroacetic acid); operating temperature: 64 ℃ C.; detector: 210nm)]。

[8- (2-aminoethyl) -6, 7-dihydro-MeBmt ] -1- [ (S) - (4-hydroxybutylthio) methyl-Sar ] -3-cyclosporin

Reacting [8- (2- (tert-butoxycarbonyl) aminoethyl) -6, 7-dihydro-MeBmt]-1- [ (S) - (4-hydroxybutylthio) methyl-Sar]-3-Cyclosporin (0.10g, 0.07mmol) was dissolved in dichloromethane (8ml) and placed in an ice-water bath. Trifluoroacetic acid (4ml) was added. The mixture was stirred at 0 ℃ for 2 hours. Another portion of dichloromethane (10ml) was added. The mixture was washed with brine (30ml), saturated sodium bicarbonate solution (30ml), dried over magnesium sulfate and evaporated under reduced pressure to give crude [8- (2-aminoethyl) -6, 7-dihydro-MeBmt]-1- [ (S) - (4-hydroxybutylthio) methyl-Sar]-3-cyclosporin [ formula: c₆₉H₁₂₈N₁₂O₁₃S; accurate quality: 1364.94, respectively; MS (M/z) 1365.75(M +1)⁺(ii) a HPLC RT: 10.57 min. (C8 reverse phase column: 250 mm; acetonitrile/water (0.05% trifluoroacetic acid); operating temperature: 64 ℃ C.; detector: 210nm)]。

[8- (2- (2-ethoxy-2-oxoethyl) amino) ethyl-6, 7-dihydro-MeBmt]-1- [ (S) - (4-hydroxybutylthio) methyl-Sar]-3-Cyclosporin (c)12a) And [8- (2-bis (2-ethoxy-2-oxoethyl) amino) Ethyl-6, 7-dihydro-MeBmt]-1- [ (S) - (4-hydroxybutylthio) methyl-Sar]-3-Cyclosporin (c)12b)

The crude [8- (2-aminoethyl) -6, 7-dihydro-MeBmt from the previous step]-1- [ (S) - (4-hydroxybutylthio) methyl-Sar]-3-Cyclosporin was dissolved in dichloromethane (10 ml). Ethyl bromoacetate (48mg, 0.34mmol) and triethylamine (0.50ml) were added. The mixture was stirred and heated to reflux for 3 hours. Dichloromethane (30ml) and brine (50ml) were then added and separated. The dichloromethane layer was dried over magnesium sulfate and evaporated under reduced pressure. The residue was purified by chromatography (dichloromethane/methanol) to give 10mg of [8- (2- ((2-ethoxy-2-oxoethyl) amino) ethyl) -6, 7-dihydro-MeBmt]-1- [ (S) - (4-hydroxybutylthio) methyl-Sar]-3-cyclosporin [ formula: c₇₃H₁₃₄N₁₂O₁₅S; accurate quality: 1450.98, respectively; MS (M/z) 1451.70(M +1)⁺,1473.87(M+Na)⁺(ii) a HPLC RT: 12.23 min. (C8 reverse phase column: 250 mm; acetonitrile/water (0.05% trifluoroacetic acid); operating temperature: 64 ℃ C.; detector: 210nm)]And 30mg of [8- (2- (bis (2-ethoxy-2-oxoethyl) amino) ethyl) -6, 7-dihydro-MeBmt]-1- [ (S) - (4-hydroxybutylthio) methyl-Sar]-3-cyclosporin [ formula: c₇₇H₁₄₀N₁₂O₁₇S; accurate quality: 1537.02, respectively; MS (M/z) 1337.70(M +1)⁺,1559.90(M+Na)⁺(ii) a HPLC RT: 14.42 min. (C8 reverse phase column: 250 mm; acetonitrile/water (0.05% trifluoroacetic acid); operating temperature: 64 ℃ C.; detector: 210nm)]。

Example 13

[8- (2- (carboxymethyl) amino) ethyl-6, 7-dihydro-MeBmt]-1- [ (S) - (4-hydroxybutylthio) methyl- Sar]-3-Cyclosporin

Reacting [8- (2- (2-ethoxy-2-oxoethyl) amino) ethyl-6, 7-dihydro-MeBmt]-1- [ (S) - (4-hydroxybutylthio) methyl-Sar]-3-Cyclosporin (10mg, 6.9X 10)^-3mmol) was dissolved in methanol (3 ml). Lithium hydroxide (2mg, 0.08mmol) and water (3ml) were added. The mixture was stirred at room temperature for one hour. Most of the methanol was then evaporated under reduced pressure. Ethyl acetate (10ml) and brine (10ml) were added, and the pH of the aqueous layer was adjusted to 3 by the addition of hydrochloric acid solution (1.0N). After separation, the ethyl acetate layer was dried over magnesium sulfate and evaporated under reduced pressure. The residue was purified by chromatography to give [8- (2- ((carboxymethyl) amino) ethyl) -6, 7-dihydro-MeBmt]-1- [ (S) - (4-hydroxybutylthio) methyl-Sar]-3-cyclosporin [ formula: c₇₁H₁₃₀N₁₂O₁₅S; accurate quality: 1422.95, respectively; MS (M/z) 1423.59(M +1)⁺,1445.84(M+Na)⁺(ii) a HPLC RT: 10.41 min. (C8 reverse phase column: 250 mm; acetonitrile/water (0.05% trifluoroacetic acid); operating temperature: 64 ℃ C.; detector: 210nm)]

Example 14

[8- (2-bis (carboxymethyl) amino) ethyl-6, 7-dihydro-MeBmt]-1- [ (S) - (4-hydroxybutylthio) methyl- Sar]-3-Cyclosporin

Reacting [8- (2-bis (2-ethoxy-2-oxoethyl) amino) ethyl-6, 7-dihydro-MeBmt]-1- [ (S) - (4-hydroxybutylthio) methyl-Sar]-3-Cyclosporin (30mg, 0.02mmol) was dissolved in methanol (3 ml). Lithium hydroxide (6mg, 0.23mmol) and water (3ml) were added. The mixture was stirred at room temperature for one hour. Most of the methanol was then evaporated under reduced pressure. Ethyl acetate (10ml) and brine (10ml) were added, and the pH of the aqueous layer was adjusted to 3 by the addition of hydrochloric acid solution (1.0N). After separation, the ethyl acetate layer was dried over magnesium sulfate and evaporated under reduced pressure. Purifying the residue by chromatography to obtain[8- (2- (bis (carboxymethyl) amino) ethyl) -6, 7-dihydro-MeBmt]-1- [ (S) - (4-hydroxybutylthio) methyl-Sar]-3-cyclosporin [ formula: c₇₃H₁₃₂N₁₂O₁₇S; accurate quality: 1480.96, respectively; MS (M/z) 1481.70(M +1)⁺,1503.87(M+Na)⁺(ii) a HPLC RT: 9.88 min. (C8 reverse phase column: 250 mm; acetonitrile/water (0.05% trifluoroacetic acid); operating temperature: 64 ℃ C.; detector: 210nm)]。

Example 15

[8- (2- (4- (methoxycarbonyl) benzyl) amino) ethyl-6, 7-dihydro-MeBmt]-1- [ (S) - (4-Hydroxybutane) Thio) methyl-Sar]-3-Cyclosporin

Reacting [8- (2-aminoethyl) -6, 7-dihydro-MeBmt]-1- [ (S) - (4-hydroxybutylthio) methyl-Sar]-3-Cyclosporin (0.50g, 0.37mmol) was dissolved in dichloromethane (50 ml). Methyl 4-formylbenzoate (0.08g, 0.46mmol), tetramethylammonium triacetoxyborohydride (0.12g, 0.46mmol) and acetic acid (5 drops) were added. The mixture was stirred at room temperature for one hour. Dichloromethane (30ml) and saturated sodium bicarbonate solution (50ml) were then added and separated. The dichloromethane layer was dried over magnesium sulfate and evaporated under reduced pressure. The residue was purified by chromatography (dichloromethane/methanol) to give 140mg of [8- (2- (4- (methoxycarbonyl) benzyl) amino) ethyl-6, 7-dihydro-MeBmt]-1- [ (S) - (4-hydroxybutylthio) methyl-Sar]-3-cyclosporin [ formula: c₇₈H₁₃₆N₁₂O₁₅S; accurate quality: 1513.00, respectively; MS (M/z) 1513.77(M +1)⁺,1535.80(M+Na)⁺(ii) a HPLC RT: 13.40 min. (C8 reverse phase column: 250 mm; acetonitrile/water (0.05% trifluoroacetic acid); operating temperature: 64 ℃ C.; detector: 210nm)]。

Example 16

[8- (2- (4-carboxybenzyl) amino) ethyl) -6, 7-dihydro-MeBmt]-1- [ (S) - (4-hydroxybutylthio) methyl base-Sar]-3-Cyclosporin

Reacting [8- (2- (4- (methoxycarbonyl) benzyl) amino) ethyl-6, 7-dihydro-MeBmt]-1- [ (S) - (4-hydroxybutylthio) methyl-Sar]-3-Cyclosporin (0.14g, 0.09mmol) was dissolved in methanol (5 ml). Lithium hydroxide (0.01g, 0.40mmol) and water (5ml) were added. The mixture was stirred at room temperature for 2 hours. Most of the methanol was then evaporated under reduced pressure. Ethyl acetate (10ml) and brine (10ml) were added, and the pH of the aqueous layer was adjusted to 3 by the addition of hydrochloric acid solution (1.00N). After separation, the ethyl acetate layer was dried over magnesium sulfate and evaporated under reduced pressure. The residue was purified by chromatography to give [8- (2- (4-carboxybenzyl) amino) ethyl-6, 7-dihydro-MeBmt]-1- [ (S) - (4-hydroxybutylthio) methyl-Sar]-3-cyclosporin [ formula: c₇₇H₁₃₄N₁₂O₁₅S; accurate quality: 1498.98, respectively; MS (M/z) 1499.62(M +1)⁺,1521.77(M+Na)⁺(ii) a HPLC RT: 11.76 min. (C8 reverse phase column: 250 mm; acetonitrile/water (0.05% trifluoroacetic acid); operating temperature: 64 ℃ C.; detector: 210nm)]。

Example 17

[8- (2- (4- (dimethylcarbamoyl)) benzyl) amino) ethyl-6, 7-dihydro-MeBmt]-1- [ (S) - (4-hydroxy) Butylthio) methyl-Sar]-3-Cyclosporin

Reacting [8- (2- (4-carboxybenzyl) amino) ethyl-6, 7-dihydro-MeBmt]-1- [ (S) - (4-hydroxybutylthio) methyl-Sar]-3-Cyclosporin (0.13g, 0.09mmol) was dissolved in dichloromethane (10 ml). Dimethylamine hydrochloride (0.04, 0.45 mm) was addedol), HBTU (0.10g, 0.27mmol), 1-hydroxybenzotriazole (0.04g, 0.27mmol) and triethylamine (0.5 ml). The mixture was stirred at room temperature for 2 hours. Dichloromethane (30ml) and brine (50ml) were then added and separated. The dichloromethane layer was dried over magnesium sulfate and evaporated under reduced pressure. The residue was purified by chromatography to give [8- (2- (4- (dimethylcarbamoyl)) benzyl) amino) ethyl-6, 7-dihydro-MeBmt]-1- [ (S) - (4-hydroxybutylthio) methyl-Sar]-3-cyclosporin [ formula: c₇₉H₁₃₉N₁₃O₁₄S; accurate quality: 1526.03, respectively; MS (M/z) 1526.72(M +1)⁺,1548.84(M+Na)⁺(ii) a HPLC RT: 11.90 min. (C8 reverse phase column: 250 mm; acetonitrile/water (0.05% trifluoroacetic acid); operating temperature: 64 ℃ C.; detector: 210nm)]。

Example 18

[8- (2-bis (4- (methoxycarbonyl) benzyl) amino)) ethyl-6, 7-dihydro-MeBmt]-1- [ (S) - (4-hydroxy) Butylthio) methyl-Sar]-3-Cyclosporin

Reacting [8- (2-aminoethyl) -6, 7-dihydro-MeBmt]-1- [ (S) - (4-hydroxybutylthio) methyl-Sar]-3-Cyclosporin (0.50g, 0.37mmol) was dissolved in dichloromethane (50 ml). Methyl 4-formylbenzoate (0.30g, 1.83mmol), tetramethylammonium triacetoxyborohydride (0.48g, 1.83mmol) and acetic acid (10 drops) were added. The mixture was stirred at room temperature for one hour. Dichloromethane (30ml) and saturated sodium bicarbonate solution (50ml) were then added and separated. The dichloromethane layer was dried over magnesium sulfate and evaporated under reduced pressure. The residue was purified by chromatography (dichloromethane/methanol) to give 174mg of [8- (2-bis (4- (methoxycarbonyl) benzyl) amino)) ethyl-6, 7-dihydro-MeBmt]-1- [ (S) - (4-hydroxybutylthio) methyl-Sar]-3-cyclosporin [ formula: c₈₇H₁₄₄N₁₂O₁₇S; accurate quality: 1661.05, respectively; MS (M/z) 1661.66(M +1)⁺,1684.06(M+Na)⁺(ii) a HPLC RT: 15.32 min. (C8 reverse phase column: 250 mm; acetonitrile/water (0.05% trifluoroacetic acid); operating temperature: 64 ℃ C.; detector: 210nm)]。

Example 19

[8- (2- (bis (4-carboxybenzyl) amino) ethyl) -6, 7-dihydro-MeBmt]-1- [ (S) - (4-hydroxybutylthio) methyl-Sar]-3-Cyclosporin

Reacting [8- (2-bis (4- (methoxycarbonyl) benzyl) amino)) ethyl-6, 7-dihydro-MeBmt]-1- [ (S) - (4-hydroxybutylthio) methyl-Sar]-3-Cyclosporin (174mg, 0.10mmol) was dissolved in methanol (5 ml). Lithium hydroxide (12mg, 0.46mmol) and water (5ml) were added. The mixture was stirred at room temperature for 2 hours. Most of the methanol was then evaporated under reduced pressure. Ethyl acetate (10ml) and brine (10ml) were added, and the pH of the aqueous layer was adjusted to 3 by the addition of hydrochloric acid solution (1.00N). After separation, the ethyl acetate layer was dried over magnesium sulfate and evaporated to give 100mg of [8- (2- (bis (4-carboxybenzyl) amino) ethyl) -6, 7-dihydro-MeBmt]-1- [ (S) - (4-hydroxybutylthio) methyl-Sar]-3-cyclosporin [ formula: c₈₅H₁₄₀N₁₂O₁₇S; accurate quality: 1633.02, respectively; MS (M/z) 1633.56(M +1)⁺,1655.68(M+Na)⁺(ii) a HPLC RT: 11.64 min. (C8 reverse phase column: 250 mm; acetonitrile/water (0.05% trifluoroacetic acid); operating temperature: 64 ℃ C.; detector: 210nm)]。

Example 20

[8- (2-bis (4- (dimethylcarbamoyl)) benzyl) amino) ethyl-6, 7-dihydro-MeBmt]-1-[(S)-(4- Hydroxybutylthio) methyl-Sar]-3-Cyclosporin

Reacting [8- (2- (bis (4-carboxybenzyl) amino) ethyl) -6, 7-dihydro-MeBmt]-1- [ (S) - (4-hydroxybutylthio) methyl-Sar]-3-Cyclosporin (0.10g, 0.06mmol) was dissolved in dichloromethane (10 ml). Dimethylamine hydrochloride (0.03, 0.37mmol), HBTU (0.12g, 0.31mmol), 1-hydroxybenzotriazole (0.04g, 0.31mmol) and triethylamine (0.5ml) were added. The mixture was stirred at room temperature for 2 hours. Dichloromethane (30ml) and brine (50ml) were then added and separated. The dichloromethane layer was dried over magnesium sulfate and evaporated under reduced pressure. The residue was purified by chromatography to give [8- (2- (bis (4- (dimethylcarbamoyl)) benzyl) amino) ethyl) -6, 7-dihydro-MeBmt]-1- [ (S) - (4-hydroxybutylthio) methyl-Sar]-3-cyclosporin [ formula: c₈₉H₁₅₀N₁₄O₁₅S; accurate quality: 1687.11, respectively; MS (M/z) 1687.73(M +1)⁺,1709.88(M+Na)⁺(ii) a HPLC RT: 12.44 min. (C8 reverse phase column: 250 mm; acetonitrile/water (0.05% trifluoroacetic acid); operating temperature: 64 ℃ C.; detector: 210nm)]。

Example 21

[8- (2- (3-Carboxybenzamido) ethyl) -6, 7-dihydro-MeBmt]-1- [ (S) - (4-hydroxybutylthio) methyl base-Sar]-3-Cyclosporin

Reacting [8- (2-aminoethyl) -6, 7-dihydro-MeBmt]-1- [ (S) - (4-hydroxybutylthio) methyl-Sar]-3-Cyclosporin (0.19g, 0.14mmol) was dissolved in dichloromethane (8 ml). Isophthalic acid (0.07g, 0.41mmol), HBTU (0.16g, 0.41mmol), 1-hydroxybenzotriazole (0.06g, 0.41mmol) and N-ethyldiisopropylamine (0.50ml) were added. The mixture was stirred at room temperature for 2 hours. Dichloromethane (50ml) and brine (50ml) were then added. The pH of the aqueous layer was adjusted to 4 by adding hydrochloric acid solution (1.0N). After separation of the mixture, the dichloromethane layer was dried over magnesium sulfate and evaporated under reduced pressureTo give crude [8- (2- (3-carboxybenzamido) ethyl) -6, 7-dihydro-MeBmt]-1- [ (S) - (4-hydroxybutylthio) methyl-Sar]-3-cyclosporin [ formula: c₇₇H₁₃₂N₁₂O₁₆S; accurate quality: 1512.96, respectively; MS (M/z) 1513.66(M +1)⁺,1535.80(M+Na)⁺(ii) a HPLC RT: 14.15 min. (C8 reverse phase column: 250 mm; acetonitrile/water (0.05% trifluoroacetic acid); operating temperature: 64 ℃ C.; detector: 210nm)]。

Crude [8- (2- (3-carboxybenzamido) ethyl) -6, 7-dihydro-MeBmt]-1- [ (S) - (4-hydroxybutylthio) methyl-Sar]-3-Cyclosporin was dissolved in acetone (10 ml). Methyl iodide (0.06g, 0.41mmol) and potassium carbonate (0.06g, 0.41mmol) were added. The mixture was stirred at room temperature for 2 hours. Most of the solvent was then evaporated. Ethyl acetate (50ml) and brine (50ml) were added and the mixture was separated. The ethyl acetate layer was dried over magnesium sulfate and evaporated under reduced pressure. The residue was purified by column (hexane/acetone) to give 44mg of [8- (2- (3- (methoxycarbonyl) benzamido) ethyl) -6, 7-dihydro-MeBmt]-1- [ (S) - (4-hydroxybutylthio) methyl-Sar]-3-cyclosporin. [ molecular formula: c₇₈H₁₃₄N₁₂O₁₆S; accurate quality: 1526.98, respectively; MS (M/z) 1527.62(M +1)⁺,1549.81(M+Na)⁺(ii) a HPLC RT: 16.08 min. (C8 reverse phase column: 250 mm; acetonitrile/water (0.05% trifluoroacetic acid); operating temperature: 64 ℃ C.; detector: 210nm)]。

Example 22

[8- (2- (3-Carboxybenzamido) ethyl) -6, 7-dihydro-MeBmt]-1- [ (S) - (4-hydroxybutylthio) methyl base-Sar]-3-Cyclosporin

Reacting [8- (2- (3- (methoxycarbonyl) benzamido) ethyl) -6, 7-dihydro-MeBmt]-1- [ (S) - (4-hydroxybutylthio) methyl-Sar]-3-Cyclosporin (44mg, 0.03mmol) dissolved in methanol (3ml). Lithium hydroxide (20mg, 0.83mmol) and water (3ml) were added. The mixture was stirred at room temperature for 2 hours. Most of the methanol was then evaporated under reduced pressure. Ethyl acetate (10ml) and brine (10ml) were added, and the pH of the aqueous layer was adjusted to 4 by the addition of hydrochloric acid solution (1.0N). After separation of the mixture, the ethyl acetate layer was dried over magnesium sulfate and evaporated under reduced pressure. The residue was purified by chromatography (C8 reverse phase column) to give 16mg of [8- (2- (3-carboxybenzamido) ethyl) -6, 7-dihydro-MeBmt]-1- [ (S) - (4-hydroxybutylthio) methyl-Sar]-3-cyclosporin [ formula: c₇₇H₁₃₂N₁₂O₁₆S; accurate quality: 1512.96, respectively; MS (M/z) 1513.66(M +1)⁺,1535.80(M+Na)⁺(ii) a HPLC RT: 14.15 min. (C8 reverse phase column: 250 mm; acetonitrile/water (0.05% trifluoroacetic acid); operating temperature: 64 ℃ C.; detector: 210nm)]。

Example 23

[8- (2- (4- (methoxycarbonyl) benzamido) ethyl) -6, 7-dihydro-MeBmt]-1- [ (S) - (4-hydroxy) Butylthio) methyl-Sar]-3-Cyclosporin

Reacting [8- (2-aminoethyl) -6, 7-dihydro-MeBmt]-1- [ (S) - (4-hydroxybutylthio) methyl-Sar]-3-Cyclosporin (0.60g, 0.44mmol) was dissolved in dichloromethane (30 ml). Monomethyl terephthalate (0.24g, 1.32mmol), HBTU (0.52g, 1.32mmol), 1-hydroxybenzotriazole (0.16g, 1.32mmol) and N, N-diisopropylethylamine (3.0ml) were added. The mixture was stirred at room temperature for 2 hours. Dichloromethane (100ml) and brine (100ml) were then added and separated. The dichloromethane layer was dried over magnesium sulfate and evaporated under reduced pressure. The residue was purified by chromatography to give 0.34g of [8- (2- (4- (methoxycarbonyl) benzamido) ethyl) -6, 7-dihydro-MeBmt]-1- [ (S) - (4-hydroxybutylthio) methyl-Sar]-3-cyclosporin [ formula: c₇₈H₁₃₄N₁₂O₁₆S; accurate quality: 1526.98, respectively; MS (m/z):1527.69(M+1)⁺,1549.95(M+Na)⁺(ii) a HPLC RT: 16.08 min. (C8 reverse phase column: 250 mm; acetonitrile/water (0.05% trifluoroacetic acid); operating temperature: 64 ℃ C.; detector: 210nm)]。

Example 24

[8- (2- (4-Carboxybenzamido) ethyl) -6, 7-dihydro-MeBmt]-1- [ (S) - (4-hydroxybutylthio) methyl base-Sar]-3-Cyclosporin

Reacting [8- (2- (4- (methoxycarbonyl) benzamido) ethyl) -6, 7-dihydro-MeBmt]-1- [ (S) - (4-hydroxybutylthio) methyl-Sar]-3-Cyclosporin (0.34mg, 0.24mmol) was dissolved in methanol (5 ml). Lithium hydroxide (20mg, 0.83mmol) and water (5ml) were added. The mixture was stirred at room temperature for 2 hours. Most of the methanol was then evaporated under reduced pressure. Ethyl acetate (20ml) and brine (20ml) were added and the pH of the aqueous layer was adjusted to 3 by the addition of hydrochloric acid solution (1.00N). After separation, the ethyl acetate layer was dried over magnesium sulfate and evaporated under reduced pressure. The residue was purified by chromatography (C18 reverse phase column) to give 0.22g of [8- (2- (4-carboxybenzamido) ethyl) -6, 7-dihydro-MeBmt]-1- [ (S) - (4-hydroxybutylthio) methyl-Sar]-3-cyclosporin [ formula: c₇₇H₁₃₂N₁₂O₁₆S; accurate quality: 1512.96, respectively; MS (M/z) 1513.66(M +1)⁺,1535.80(M+Na)⁺(ii) a HPLC RT: 13.98 min. (C8 reverse phase column: 250 mm; acetonitrile/water (0.05% trifluoroacetic acid); operating temperature: 64 ℃ C.; detector: 210nm)]。

Example 25

[8- (2- (4- (dimethylcarbamoyl) benzamido) ethyl) -6, 7-dihydro-MeBmt]-1-[(S)-(4- Hydroxybutylthio) methyl-Sar]-3-Cyclosporin

Reacting [8- (2- (4-carboxybenzamido) ethyl) -6, 7-dihydro-MeBmt]-1- [ (S) - (4-hydroxybutylthio) methyl-Sar]-3-Cyclosporin (0.22g, 0.15mmol) was dissolved in dichloromethane (5 ml). Dimethylamine hydrochloride (0.04, 0.49mmol), HBTU (0.17g, 0.45mmol), 1-hydroxybenzotriazole (0.06g, 0.45mmol) and pyridine (0.5ml) were added. The mixture was stirred at room temperature overnight. The dichloromethane was then evaporated under reduced pressure. Ethyl acetate (50ml) and brine (50ml) were added and separated. The dichloromethane layer was dried over magnesium sulfate and evaporated under reduced pressure. The residue was purified by chromatography to give [8- (2- (4- (dimethylcarbamoyl) benzamido) ethyl) -6, 7-dihydro-MeBmt]-1- [ (S) - (4-hydroxybutylthio) methyl-Sar]-3-cyclosporin [ formula: c₇₉H₁₃₇N₁₃O₁₅S; accurate quality: 1540.01, respectively; MS (M/z) 1540.89(M +1)⁺,1563.06(M+Na)⁺(ii) a HPLC RT: 14.54 min. (C8 reverse phase column: 250 mm; acetonitrile/water (0.05% trifluoroacetic acid); operating temperature: 64 ℃ C.; detector: 210nm)]。

Example 26

[8- (2- (2-Hydroxyacetylamino) ethyl) -6, 7-dihydro-MeBmt]-1- [ (S) - (4-hydroxybutylthio) methyl base-Sar]-3-Cyclosporin

Reacting [8- (2-aminoethyl) -6, 7-dihydro-MeBmt]-1- [ (S) - (4-hydroxybutylthio) methyl-Sar]-3-Cyclosporin (0.26g, 0.19mmol) was dissolved in dichloromethane (20 ml). Glycolic acid (0.04g, 0.53mmol), HBTU (0.22g, 0.58mmol), 1-hydroxybenzotriazole (0.08g, 0.58mmol) and pyridine (1.00ml) were added. The mixture was stirred at room temperature for 2 hours. Dichloromethane (30ml) and brine (50ml) were then added and separated. Drying of dichloro-benzene over magnesium sulfateThe methane layer was evaporated under reduced pressure. The residue was purified by chromatography (dichloromethane/methanol) to give pure [8- (2- (2-hydroxyacetamido) ethyl) -6, 7-dihydro-MeBmt]-1- [ (S) - (4-hydroxybutylthio) methyl-Sar]-3-cyclosporin [ formula: c₇₁H₁₃₀N₁₂O₁₅S; accurate quality: 1422.95, respectively; MS (M/z) 1423.80(M +1)⁺,1445.95(M+Na)⁺(ii) a HPLC RT: 12.73 min. (C8 reverse phase column: 250 mm; acetonitrile/water (0.05% trifluoroacetic acid); operating temperature: 64 ℃ C.; detector: 210nm)]。

Example 27

[8- (2-Acetylaminoethyl) -6, 7-dihydro-MeBmt]-1- [ (R) - (4-acetoxybutoxy) methyl- Sar]-3-Cyclosporin (26a)

Reacting [8- (2-acetamido ethyl) -6, 7-dihydro-MeBmt]-1- [ (R) -hydroxymethyl-Sar]-3-Cyclosporin (0.14g, 0.11mmol) was dissolved in benzene (10 ml). 4-Bromobutyl acetate (0.21g, 1.06mmol), tetra-n-butylammonium bromide (0.34g, 1.06mmol), tetramethylammonium hydroxide pentahydrate (0.19g, 1.06mmol) and sodium hydroxide solution (2.0ml, 45%) were added. The mixture was stirred at 50 ℃ overnight. Ethyl acetate (50ml) and brine (50ml) were then added and separated. The ethyl acetate layer was dried over magnesium sulfate and evaporated under reduced pressure. The residue was purified by chromatography (dichloromethane/methanol) to give 50mg of pure [8- (2-acetamidomethyl) -6, 7-dihydro-MeBmt]-1- [ (R) - (4-acetoxybutoxy) methyl-Sar]-3-cyclosporin [ formula: c₇₃H₁₃₂N₁₂O₁₆(ii) a Accurate quality: 1432.99, respectively; MS (M/z) 1433.73(M +1)⁺,1455.99(M+Na)⁺(ii) a HPLC RT: 17.43 min. (C8 reverse phase column: 250 mm; acetonitrile/water (0.05% trifluoroacetic acid); operating temperature: 64 ℃ C.; detector: 210nm)]。

Example 28

[8- (2-Acetylaminoethyl) -6, 7-dihydro-MeBmt]-1- [ (R) - (4-Hydroxybutoxy) methyl-Sar ]-3- Cyclosporin

Reacting [8- (2-acetamido ethyl) -6, 7-dihydro-MeBmt]-1- [ (R) - (4-acetoxybutoxy) methyl-Sar]-3-Cyclosporin (50mg, 0.03mmol) was dissolved in methanol (5 ml). Lithium hydroxide (20mg, 0.83mmol) and water (5ml) were added. The mixture was stirred at room temperature for 2 hours. Most of the methanol was then evaporated under reduced pressure. Ethyl acetate (20ml) and brine (30ml) were added, and the pH of the aqueous layer was adjusted to 3 by the addition of hydrochloric acid solution (1.00N). After separation, the ethyl acetate layer was dried over magnesium sulfate and evaporated under reduced pressure. The residue was purified by C18 reverse phase chromatography to give 15mg of [8- (2-acetamidomethyl) -6, 7-dihydro-MeBmt]-1- [ (R) - (4-Hydroxybutoxy) methyl-Sar]-3-cyclosporin [ formula: c₇₁H₁₃₀N₁₂O₁₅(ii) a Accurate quality: 1390.98, respectively; MS (M/z) 1391.80(M +1)⁺,1413.98(M+Na)⁺(ii) a HPLC RT: 15.50 min. (C8 reverse phase column: 250 mm; acetonitrile/water (0.05% trifluoroacetic acid); operating temperature: 64 ℃ C.; detector: 210nm)]。

Example 29

[8- (3-Acetylaminopropyl) -6, 7-dihydro-MeBmt]-1- [ (S) - (3- (N-morpholinyl) propylthio) methyl- Sar]-3-Cyclosporin

(4-Cyanobutyl) triphenylphosphine bromide

5-Bromopentanenitrile (1.62g, 10.00mmol) and triphenylphosphine (2.62g, 10.00mmol)) Was added to toluene (20 ml). The mixture was stirred and heated to reflux overnight. After cooling to room temperature, the precipitate was filtered off, washed with toluene and hexane and dried in vacuo to give 3.00g of product. [ molecular formula: c₂₃H₂₃BrNP; accurate quality: 423.08, respectively; MS (M/z) 344.28(M-Br)⁺]。

[8- (2-cyanoethyl) -3-acetyl-MeBmt ] -1-cyclosporin

To a solution of (4-cyanobutyl) triphenyl phosphine bromide (6.40g, 15.08mmol) in anhydrous tetrahydrofuran (180ml) under nitrogen was added sodium bis (trimethylsilyl) amide (2.0M in THF, 11.2ml, 22.40 mmol). The reaction mixture was stirred at room temperature for one hour and cooled to-30 ℃. Adding [ (3R,4R) -3-acetoxy-4-methyl-6-oxo-N-Menle]-1-Cyclosporin (8.20g, 6.65mmol) in anhydrous tetrahydrofuran (25 ml). The mixture was stirred at-30 ℃ for a further 2 hours. Saturated ammonium chloride solution (20ml) was then added to quench the reaction. Most of the tetrahydrofuran was evaporated under reduced pressure. Ethyl acetate (250ml) and brine (100ml) were added and the mixture was separated. The organic layer was dried over magnesium sulfate and evaporated under reduced pressure. The residue was purified by chromatography (dichloromethane/methanol) to give 5.60g of pure [8- (2-cyanoethyl) -3-acetyl-MeBmt]-1-cyclosporin [ formula: c₆₇H₁₁₆N₁₂O₁₃(ii) a Accurate quality: 1296.88, respectively; MS (M/z) 1297.57(M +1)⁺]。

[8- (2-cyanoethyl) -MeBmt ] -1-cyclosporin

Reacting [8- (2-cyanoethyl) -3-acetyl-MeBmt]-1-Cyclosporin (5.00g, 3.86mmol) was dissolved in methanol (100 ml). Water (50ml) and tetramethylammonium hydroxide pentahydrate (4.40g, 24.22mmol) were added. The mixture was stirred at room temperature overnight. Most of the methanol is then evaporated. Ethyl acetate (250ml) and brine (100ml) were added and the mixture was separated. The organic layer was dried over magnesium sulfate and evaporated under reduced pressure. The residue was purified by chromatography (dichloromethane/methanol) to give 3.60g of pure [8- (2-cyanoethyl) -MeBmt]-1-cyclosporin [ formula: c₆₅H₁₁₄N₁₂O₁₂(ii) a Accurate quality: 1254.87, respectively; MS (M/z) 1255.51(M +1)⁺(ii) a HPLC RT: 14.90 min. (C8 reverse phase column: 250 mm; acetonitrile/water (0.05% trifluoroacetic acid); operating temperature: 64 ℃ C.; detector: 210nm)]。

[8- (3-aminopropyl) -6, 7-dihydro-MeBmt ] -1-cyclosporin

To [8- (2-cyanoethyl) -MeBmt under nitrogen]To a solution of (2.40g, 1.91mmol) of (E) -1-cyclosporin in methanol (120ml) was added nickel (II) chloride hexahydrate (0.38g, 1.62 mmol). Sodium borohydride (2.00g, 52.63mmol) was added in four portions over 30 minutes. After stirring the mixture for a further 1 hour, most of the methanol was evaporated under reduced pressure. Ethyl acetate (100ml) and saturated sodium bicarbonate solution (50ml) were added to the solution, and the mixture was separated. The organic layer was washed with brine, dried over magnesium sulfate and evaporated under reduced pressure. The residue was dissolved in methanol (30 ml). Palladium (10 wt% on carbon, 150mg) and acetic acid (5 drops) were added. The mixture was stirred at room temperature under hydrogen for 4 hours. The mixture was filtered and the filtrate was evaporated under reduced pressure to give crude [8- (3-aminopropyl) -6, 7-dihydro-MeBmt]-1-cyclosporin [ formula: c₆₅H₁₂₀N₁₂O₁₂(ii) a Accurate quality: 1260.90, respectively; MS (M/z) 1261.69(M +1)⁺(ii) a HPLC RT: 11.23 min. (C8 reverse phase column: 250 mm; acetonitrile/water (0.05% trifluoroacetic acid); operating temperature: 64 ℃ C.; detector: 210nm)]。

[8- (3-Acetylaminopropyl) -6, 7-dihydro-MeBmt ] -1-cyclosporin

To [8- (3-aminopropyl) -6, 7-dihydro-MeBmt]To a solution of-1-cyclosporin (2.40g, 1.90mmol) and acetic acid (240mg, 60mmol) in dichloromethane (100ml) were added diisopropylethylamine (386mg, 1.99mmol) and HATU (1.20g, 3.15 mmol). After stirring at room temperature for 2 hours, the reaction mixture was washed with saturated sodium bicarbonate solution, brine, dried over magnesium sulfate and evaporated under reduced pressure. The residue was purified by chromatography (dichloromethane/methanol) to give 2.00g of pure [8- (3-acetamidopropyl) -6, 7-dihydro-MeBmt]-1-cyclosporin [ formula: c₆₇H₁₂₂N₁₂O₁₃(ii) a Accurate quality: 1302.93, respectively; MS (M/z) 1303.63(M +1)⁺. HPLC RT:13.74 min. (C8 reverse phase column: 250 mm; acetonitrile/water (0.05% trifluoroacetic acid); operating temperature: 64 ℃ C.; detector: 210nm)]。

[8- (3-Acetylaminopropyl) -6, 7-dihydro-MeBmt ] -1- [ α -methylene-Sar ] -3-cyclosporin

N-butyllithium (2.65M,12ml, 31.80mmol) was added to a solution of diisopropylamine (3.23g, 32mmol) in tetrahydrofuran (100ml) at-78 ℃ under nitrogen. After stirring the reaction mixture for 1 hour, [8- (3-acetamidopropyl) -6, 7-dihydro-MeBmt ] was added over 10 minutes]-1-Cyclosporin (3.80g, 2.92mmol) in tetrahydrofuran (15 ml). The mixture was stirred at-78 ℃ for three hours. After bubbling carbon dioxide gas into the reaction mixture for 30 minutes, the mixture was stirred at-78 ℃ for another 1 hour. The cooling bath was then removed and the reaction mixture was allowed to warm to room temperature to allow unreacted carbon dioxide to vent. The mixture was cooled to-78 ℃ and chloromethyl chloroformate (3.70ml) was added. The mixture was stirred and allowed to warmWarm to room temperature overnight. Brine (20ml) was added to quench the reaction. Most of the tetrahydrofuran was removed under reduced pressure. Ethyl acetate (50ml) and brine (50ml) were added and the mixture was separated. The organic layer was dried over magnesium sulfate and evaporated under reduced pressure. The residue was purified by chromatography (hexane/acetone) to give 1.20g of pure [8- (3-acetamidopropyl) -6, 7-dihydro-MeBmt]-1- [ α -methylene-Sar]-3-cyclosporin [ formula: c₆₈H₁₂₂N₁₂O₁₃(ii) a Accurate quality: 1314.93, respectively; MS (M/z) 1315.61(M +1)⁺(ii) a HPLCRT: 16.44 min. (C8 reverse phase column: 250 mm; acetonitrile/water (0.05% trifluoroacetic acid); operating temperature: 64 ℃ C.; detector: 210nm)]。

[8- (3-Acetylaminopropyl) -6, 7-dihydro-MeBmt ] -1- [ (S) - (3- (N-morpholinyl) propylthio) methyl-Sar ] -3-cyclosporin

To [8- (3-acetamidopropyl) -6, 7-dihydro-MeBmt]-1- [ (S) - (α -methylene-Sar)]To a solution of (0.50g, 0.38mmol) of (E) -3-cyclosporin in methanol (60ml) were added 3-morpholinopropanethiol (0.48g, 3.00mmol) and lithium hydroxide (92mg, 3.83 mmol). The reaction mixture was stirred at room temperature overnight. Most of the methanol was evaporated under reduced pressure. Ethyl acetate (80ml) and brine (30ml) were added to the solution, and the mixture was separated. The organic layer was dried over magnesium sulfate and evaporated under reduced pressure. The residue was purified by chromatography (dichloromethane/methanol) to give 130mg of pure [8- (3-acetamidopropyl) -6, 7-dihydro-MeBmt]-1- [ (S) - (3- (N-morpholinyl) propylthio) methyl-Sar]-3-cyclosporin [ formula: c₇₅H₁₃₇N₁₃O₁₄S; accurate quality: 1476.01, respectively; MS (M/z) 1476.82(M +1)⁺(ii) a HPLC RT: 12.61 min. (C8 reverse phase column: 250 mm; acetonitrile/water (0.05% trifluoroacetic acid); operating temperature: 64 ℃ C.; detector: 210nm)]。

Example 30

[8- (3-Acetylaminopropan)Yl) -6, 7-dihydro-MeBmt]-1- [ (S) - (4-hydroxybutylthio) methyl-Sar]-3- Cyclosporin

To [8- (3-acetamidopropyl) -6, 7-dihydro-MeBmt]-1- [ (S) - (α -methylene-Sar)]To a solution of (0.50g, 0.38mmol) of (E) -3-cyclosporin in methanol (60ml) were added (0.28g, 2.64mmol) of 4-mercapto-1-butanol and (92mg, 3.83mmol) of lithium hydroxide. The reaction mixture was stirred at room temperature overnight. Most of the methanol was evaporated under reduced pressure. Ethyl acetate (80ml) and brine (30ml) were added to the solution, and the mixture was separated. The organic layer was dried over magnesium sulfate and evaporated under reduced pressure. The residue was purified by chromatography (dichloromethane/methanol) to give 115mg of pure [8- (3-acetamidopropyl) -6, 7-dihydro-MeBmt]-1- [ (S) - (4-hydroxybutylthio) methyl-Sar]-3-cyclosporin [ formula: c₇₂H₁₃₂N₁₂O₁₄S; accurate quality: 1420.97, respectively; MS (M/z) 1421.80(M +1)⁺(ii) a HPLC RT: 15.49 min. (C8 reverse phase column: 250 mm; acetonitrile/water (0.05% trifluoroacetic acid); operating temperature: 64 ℃ C.; detector: 210nm)]。

Example 31

[8- (3- (4-Carboxybenzamido) propyl) -6, 7-dihydro-MeBmt]-1- [ (S) - (4-hydroxybutylthio) methyl base-Sar]-3-Cyclosporin

[8- (3- (4-Methoxycarbonylbenzamido) propyl) -6, 7-dihydro-MeBmt ] -1-cyclosporin

To [8- (3-aminopropyl) -6, 7-dihydro-MeBmt](ii) -1-Cyclosporin (4.00g, 3.17mmol) and monomethyl terephthalate (0.85g, 4.72mmol) in dichloromethane (100ml) were added diisopropylethylamine (770mg, 5.96mmol) and HATU (2.40g, 6.30 mmol). The reaction mixture was stirred at room temperature for 2 hours, then washed with saturated sodium bicarbonate solution, brine, dried over magnesium sulfate and evaporated under reduced pressure. The residue was purified by chromatography (dichloromethane/methanol) to give 2.00g of pure [8- (3- (4-methoxycarbonylbenzamido) propyl) -6, 7-dihydro-MeBmt]-1-cyclosporin [ formula: c₇₄H₁₂₆N₁₂O₁₅(ii) a Accurate quality: 1422.95, respectively; MS (M/z) 1423.70(M +1)⁺. HPLC RT 16.74 min. (C8 reverse phase column: 250 mm; acetonitrile/water (0.05% trifluoroacetic acid); operating temperature: 64 ℃ C.; detector: 210nm)]。

[8- (3- (4-Carboxybenzamido) propyl) -6, 7-dihydro-MeBmt ] -1-Cyclosporin

To [8- (3- (4-methoxycarbonylbenzamido) propyl) -6, 7-dihydro-MeBmt]To a solution of-1-cyclosporin (2.40g, 1.68mmol) in methanol (80ml) and water (20ml) was added lithium hydroxide (0.28g, 11.66 mmol). The reaction mixture was stirred at room temperature for 2 hours and evaporated under reduced pressure. Ethyl acetate (10ml) and brine (10ml) were added, and the pH of the aqueous layer was adjusted to 5 by the addition of hydrochloric acid solution (1.00N). After separation, the ethyl acetate layer was dried over magnesium sulfate and evaporated to give 2.3g of [8- (3- (4-carboxybenzamido) propyl) -6, 7-dihydro-MeBmt]-1-cyclosporin. [ molecular formula: c₇₃H₁₂₄N₁₂O₁₅(ii) a Accurate quality: 1408.93, respectively; MS (M/z) 1409.70(M +1)⁺. HPLC RT:14.93 min. (C8 reverse phase column: 250 mm; acetonitrile/water (0.05% trifluoroacetic acid); operating temperature: 64 ℃ C.; detector: 210nm)]。

[8- (3- (4-Carboxybenzamido) propyl) -6, 7-dihydro-MeBmt ] -1- [ α -methylene-Sar ] -3-Cyclosporin

N-butyllithium (2.65M,7.6ml, 20.14mmol) was added to a solution of diisopropylamine (2.02g, 20.00mmol) in tetrahydrofuran (120ml) at-78 ℃ under nitrogen. After stirring the reaction mixture for 1 hour, add [8- (3- (4-carboxybenzamido) propyl) -6, 7-dihydro-MeBmt over 10 minutes]-1-Cyclosporin (2.10g, 1.49mmol) in tetrahydrofuran (15 ml). The mixture was stirred at-78 ℃ for three hours. After bubbling carbon dioxide gas into the reaction mixture for 30 minutes, the mixture was stirred at-78 ℃ for another 1 hour. The cooling bath was then removed and the reaction mixture was allowed to warm to room temperature to allow unreacted carbon dioxide to vent. The mixture was cooled to-78 ℃ and chloromethyl chloroformate (2.20ml) was added. The mixture was stirred and allowed to warm to room temperature overnight. Brine (20ml) was added to quench the reaction. Most of the tetrahydrofuran was removed under reduced pressure. Ethyl acetate (50ml) and brine (50ml) were added and the mixture was separated. The organic layer was dried over magnesium sulfate and evaporated under reduced pressure. The residue was purified by chromatography to give 1.20g of pure [8- (3- (4-carboxybenzamido) propyl) -6, 7-dihydro-MeBmt]-1- [ α -methylene-Sar]-3-cyclosporin [ formula: c₇₄H₁₂₄N₁₂O₁₅(ii) a Accurate quality: 1420.93, respectively; MS (M/z) 1421.61(M +1)⁺(ii) a HPLC RT: 16.56 min. (C8 reverse phase column: 250 mm; acetonitrile/water (0.05% trifluoroacetic acid); operating temperature: 64 ℃ C.; detector: 210nm)]。

[8- (3- (4-Carboxybenzamido) propyl) -6, 7-dihydro-MeBmt ] -1- [ (S) - (4-Hydroxybutylthio) methyl-Sar ] -3-Cyclosporin

To [8- (3- (4-carboxybenzamido) propyl) -6, 7-dihydro-MeBmt]-1- [ α -methylene-Sar]To a solution of (0.50g, 0.35mmol) of (E) -3-cyclosporin in methanol (30ml) were added (0.28g, 2.64mmol) of 4-mercapto-1-butanol and (92mg, 3.83mmol) of lithium hydroxide. The reaction mixture was stirred at room temperature overnight. Most of the methanol was evaporated under reduced pressure. Ethyl acetate (80ml) and brine (30ml) were added to the solution, and the mixture was separated.The organic layer was dried over magnesium sulfate and evaporated under reduced pressure. The residue was purified by chromatography (dichloromethane/methanol) to give 115mg of pure [8- (3- (4-carboxybenzamido) propyl) -6, 7-dihydro-MeBmt]-1- [ (S) - (4-hydroxybutylthio) methyl-Sar]-3-cyclosporin [ formula: c₇₈H₁₃₄N₁₂O₁₆S; accurate quality: 1526.98, respectively; MS (M/z) 1527.84(M +1)⁺(ii) a HPLC RT: 14.80 min. (C8 reverse phase column: 250 mm; acetonitrile/water (0.05% trifluoroacetic acid); operating temperature: 64 ℃ C.; detector: 210nm)]。

Example 32

[8- (3- (4- (diethylcarbamoylbenzamido) propyl) -6, 7-dihydro-MeBmt]-1- [ (S) - (4-hydroxy) Butylthio) methyl-Sar]-3-Cyclosporin

To [8- (3- (4-carboxybenzamido) propyl) -6, 7-dihydro-MeBmt]-1- [ (S) - (4-hydroxybutylthio) methyl-Sar]To a solution of (0.10g, 0.07mmol) of (E) -3-cyclosporin and (15mg, 0.21mmol) of diethylamine in dichloromethane (10ml) were added diisopropylethylamine (25mg, 0.19mmol), HOBT (29mg, 0.19mmol) and HATU (73mg, 0.19 mmol). After stirring at room temperature for 2 hours, the reaction mixture was washed with saturated sodium bicarbonate solution and brine, dried over magnesium sulfate and evaporated under reduced pressure. The residue was purified by chromatography (dichloromethane/methanol) to give 42mg of pure [8- (3- (4-diethylcarbamoylbenzamido) propyl) -6, 7-dihydro-MeBmt]-1- [ (S) - (4-hydroxybutylthio) methyl-Sar]-3-cyclosporin [ formula: c₈₂H₁₄₃N₁₃O₁₅S; accurate quality: 1582.05, respectively; MS (M/z) 1582.70(M +1)⁺. HPLC RT 16.08 min. (C8 reverse phase column: 250 mm; acetonitrile/water (0.05% trifluoroacetic acid); operating temperature: 64 ℃ C.; detector: 210nm)]。

Example 33

[8- (3- (3-Methoxycarbonylbenzamido) propyl) -6, 7-dihydro-MeBmt]-1- [ (S) - (4-Hydroxybutane) Thio) methyl-Sar]-3-Cyclosporin

[8- (3- (3-methoxycarbonylbenzamido) propyl) -6, 7-dihydro-MeBmt ] -1-cyclosporin

To [8- (3-aminopropyl) -6, 7-dihydro-MeBmt]To a solution of (4.33g, 3.43mmol) of (E) -1-cyclosporin and (0.92g, 5.11mmol) of monomethyl isophthalate in dichloromethane (100ml) were added diisopropylethylamine (770mg, 5.96mmol) and HATU (2.40g, 6.30 mmol). The reaction mixture was stirred at room temperature for 2 hours, then washed with saturated sodium bicarbonate solution, brine, dried over magnesium sulfate and evaporated under reduced pressure. The residue was purified by chromatography (dichloromethane/methanol) to give 2.00g of pure [8- (3- (3-methoxycarbonylbenzamido) propyl) -6, 7-dihydro-MeBmt]-1-cyclosporin [ formula: c₇₄H₁₂₆N₁₂O₁₅(ii) a Accurate quality: 1422.95, respectively; MS (M/z) 1423.70(M +1)⁺. HPLC RT 16.82 min. (C8 reverse phase column: 250 mm; acetonitrile/water (0.05% trifluoroacetic acid); operating temperature: 64 ℃ C.; detector: 210nm)]。

[8- (3- (3-Carboxybenzamido) propyl) -6, 7-dihydro-MeBmt ] -1-Cyclosporin

To [8- (3- (3-methoxycarbonylbenzamido) propyl) -6, 7-dihydro-MeBmt]To a solution of (2.50g, 1.76mmol) of (E) -1-cyclosporin in methanol (80ml) were added water (20ml) and lithium hydroxide (0.28g, 11.66 mmol). The reaction mixture was stirred at room temperature for 2 hours and evaporated under reduced pressure. Ethyl acetate (10ml) and brine (10ml) were added, and the pH of the aqueous layer was adjusted by adding hydrochloric acid solution (1.00N)Adjusted to 5. After separation, the ethyl acetate layer was dried over magnesium sulfate and evaporated to give 2.1g of [8- (3- (3-carboxybenzamido) propyl) -6, 7-dihydro-MeBmt]-1-cyclosporin. [ molecular formula: c₇₃H₁₂₄N₁₂O₁₅(ii) a Accurate quality: 1408.93, respectively; MS (M/z) 1409.70(M +1)⁺. HPLC RT:15.11 min. (C8 reverse phase column: 250 mm; acetonitrile/water (0.05% trifluoroacetic acid); operating temperature: 64 ℃ C.; detector: 210nm)]。

[8- (3- (3-methoxycarbonylbenzamido) propyl) -6, 7-dihydro-MeBmt ] -1- [ α -methylene-Sar ] -3-Cyclosporin

N-butyllithium (2.65M, 7.9ml, 20.94mmol) was added to a solution of diisopropylamine (2.12g, 21.00mmol) in tetrahydrofuran (120ml) at-78 ℃ under nitrogen. After stirring the reaction mixture for 1 hour, add [8- (3- (3-carboxybenzamido) propyl) -6, 7-dihydro-MeBmt over 10 minutes]-1-Cyclosporin (2.10g, 1.49mmol) in tetrahydrofuran (15 ml). The mixture was stirred at-78 ℃ for three hours. After bubbling carbon dioxide gas into the reaction mixture for 30 minutes, the mixture was stirred at-78 ℃ for another 1 hour. The cooling bath was then removed and the reaction mixture was allowed to warm to room temperature to allow unreacted carbon dioxide to vent. The mixture was cooled to-78 ℃ and chloromethyl chloroformate (2.20ml) was added. The mixture was stirred and allowed to warm to room temperature overnight. Methanol (20ml) was added to quench the reaction. Most of the tetrahydrofuran was removed under reduced pressure. Ethyl acetate (50ml) and brine (50ml) were added and the mixture was separated. The organic layer was dried over magnesium sulfate and evaporated under reduced pressure. The residue was purified by chromatography (hexane/acetone) to give 1.20g of pure [8- (3- (3-methoxycarbonylbenzamido) propyl) -6, 7-dihydro-MeBmt]-1- [ α -methylene-Sar]-3-cyclosporin [ formula: c₇₅H₁₂₆N₁₂O₁₅(ii) a Accurate quality: 1434.95, respectively; MS (M/z) 1435.61(M +1)⁺(ii) a HPLC RT: and 18.26 min. (C8 reverse phase column: 250 mm; acetonitrile/water (0.05% trifluoroacetic acid); operationWorking temperature: 64 ℃; a detector: 210nm)]。

[8- (3- (3-methoxycarbonylbenzamido) propyl) -6, 7-dihydro-MeBmt ] -1- [ (S) - (4-hydroxybutylthio) methyl-Sar ] -3-cyclosporin

To [8- (3- (3-methoxycarbonylbenzamido) propyl) -6, 7-dihydro-MeBmt]-1- [ α -methylene-Sar]To a solution of (0.62g, 0.43mmol) of (E) -3-cyclosporin in methanol (30ml) were added (0.28g, 2.64mmol) of 4-mercapto-1-butanol and (103mg, 4.29mmol) of lithium hydroxide. The reaction mixture was stirred at room temperature overnight. Most of the methanol was evaporated under reduced pressure. Ethyl acetate (80ml) and brine (30ml) were added to the solution, and the mixture was separated. The organic layer was dried over magnesium sulfate and evaporated under reduced pressure. The residue was purified by chromatography (dichloromethane/methanol) to give 115mg of pure [8- (3- (3-methoxycarbonylbenzamido) propyl) -6, 7-dihydro-MeBmt]-1- [ (S) - (4-hydroxybutylthio) methyl-Sar]-3-cyclosporin [ formula: c₇₉H₁₃₆N₁₂O₁₆S; accurate quality: 1540.99, respectively; MS (M/z) 1541.84(M +1)⁺(ii) a HPLC RT: 16.83 min. (C8 reverse phase column: 250 mm; acetonitrile/water (0.05% trifluoroacetic acid); operating temperature: 64 ℃ C.; detector: 210nm)]。

Example 34

[8- (3- (3-Carboxybenzamido) propyl) -6, 7-dihydro-MeBmt]-1- [ (S) - (4-hydroxybutylthio) methyl base-Sar]-3-Cyclosporin

To [8- (3- (3-methoxycarbonylbenzamido) propyl) -6, 7-dihydro-MeBmt]-1- [ (S) - (4-hydroxybutylthio) methyl-Sar]-3-Cyclosporin (0.20g, 0.13mmol) in methanol (8ml) was addedWater (3ml) and lithium hydroxide (15mg, 0.63 mmol). The reaction mixture was stirred at room temperature for 8 hours and evaporated under reduced pressure. Ethyl acetate (10ml) and brine (10ml) were added, and the pH of the aqueous layer was adjusted to 5 by the addition of hydrochloric acid solution (1.00N). After separation, the ethyl acetate layer was dried over magnesium sulfate and evaporated to give [8- (3- (3-carboxybenzamido) propyl) -6, 7-dihydro-MeBmt]-1- [ (S) - (4-hydroxybutylthio) methyl-Sar]-3-cyclosporin [ formula: c₇₈H₁₃₄N₁₂O₁₆S; accurate quality: 1526.98, respectively; MS (M/z) 1527.84(M +1)⁺(ii) a HPLC RT: 14.99 min. (C8 reverse phase column: 250 mm; acetonitrile/water (0.05% trifluoroacetic acid); operating temperature: 64 ℃ C.; detector: 210nm)]。

Example 35

[8- (4-Acylaminobutyl) -6, 7-dihydro-MeBmt]-1- [ (S) - (4-hydroxybutylthio) methyl-Sar]-3- Cyclosporin

(5-Cyanopentyl) triphenylphosphine bromide

6-bromohexanenitrile (10.00g, 56.80mmol) and triphenylphosphine (14.90g, 56.80mmol) were dissolved in toluene (100 ml). The mixture was stirred and heated to reflux for 3 days. After cooling to room temperature, most of the toluene was decanted off. The residue was dried under vacuum for 6 hours. Hexane (160ml) was then added and the mixture was stirred at room temperature over the weekend. The precipitate was filtered off and dried in vacuo to give 21.0g of product. [ molecular formula: c₂₄H₂₅NP⁺(ii) a Accurate quality: 358.17, respectively; MS (M/z) 358.20(M)⁺]。

[8- (3-cyanopropyl) -3-acetyl-MeBmt ] -1-cyclosporin

To a dry flask were added (5-cyanopentyl) triphenylphosphonium bromide (14.00g, 32.04mmol) and anhydrous tetrahydrofuran (50ml) under nitrogen. The reaction mixture was placed in an ice-water bath and sodium tert-butoxide (4.30g, 44.84mmol) was added. After stirring the mixture for 3 hours [ (3R,4R) -3-acetoxy-4-methyl-6-oxo-N-Menle was added]-a solution of 1-cyclosporin (7.90g, 6.40mmol) in anhydrous tetrahydrofuran (20 ml). The mixture was stirred at 0 ℃ for a further 3 hours. Saturated ammonium chloride solution (20ml) was then added to quench the reaction. Most of the tetrahydrofuran was evaporated under reduced pressure. Ethyl acetate (100ml) and brine (100ml) were added and the mixture was separated. The organic layer was dried over magnesium sulfate and evaporated under reduced pressure. The residue was purified by chromatography (dichloromethane/methanol) to give 7.19g of [8- (3-cyanopropyl) -3-acetyl-MeBmt]-1-cyclosporin [ formula: c₆₈H₁₁₈N₁₂O₁₃(ii) a Accurate quality: 1310.89, respectively; MS (M/z) 1311.58(M +1)⁺,1333.74(M+Na)⁺(ii) a HPLC RT: 17.19 min. (C8 reverse phase column: 250 mm; acetonitrile/water (0.05% trifluoroacetic acid); operating temperature: 64 ℃ C.; detector: 210nm)]。

[8- (3-cyanopropyl) -MeBmt ] -1-cyclosporin

Reacting [8- (3-cyanopropyl) -3-acetyl-MeBmt]-1-Cyclosporin (7.19g, 5.48mmol) was dissolved in methanol (40 ml). Water (20ml) and tetramethylammonium hydroxide pentahydrate (2.98g, 16.45mmol) were added. The mixture was stirred at room temperature for 2 hours. Most of the methanol is then evaporated. Ethyl acetate (100ml) and brine (100ml) were added and the mixture was separated. The organic layer was dried over magnesium sulfate and evaporated under reduced pressure. The residue was purified by chromatography (dichloromethane/methanol) to give 4.50g of pure [8- (3-cyanopropyl) -MeBmt]-1-cyclosporin [ formula: c₆₆H₁₁₆N₁₂O₁₂(ii) a Accurate quality: 1268.88, respectively; MS (M/z) 1269.62(M +1)⁺,1291.76(M+Na)⁺；HPLC RT：15.78min. (C8 reverse phase column: 250 mm; acetonitrile/water (0.05% trifluoroacetic acid); operating temperature: 64 ℃ C.; detector: 210nm)]。

[8- (4-Aminobutyl) -6, 7-dihydro-MeBmt ] -1-Cyclosporin

To [8- (3-cyanopropyl) -MeBmt under nitrogen]To a solution of (4.50g, 3.63mmol) of (E) -1-cyclosporin in methanol (60ml) was added nickel (II) chloride hexahydrate (0.43g, 1.81 mmol). The reaction mixture was placed in an ice-water bath. Sodium borohydride (7.04g, 181.33mmol) was added in four portions over a two hour period. After the mixture was stirred at 0 ℃ for another 2 hours, water (10ml) was added. Most of the methanol was evaporated under reduced pressure. Ethyl acetate (120ml) and saturated sodium bicarbonate solution (120ml) were added to the solution, and the mixture was separated. The organic layer was washed with brine, dried over magnesium sulfate and evaporated under reduced pressure. The residue was dissolved in methanol (60 ml). Palladium (10 wt% on carbon, 50mg) and acetic acid (6 drops) were added. The mixture was stirred at room temperature under hydrogen overnight. The mixture was filtered and the filtrate evaporated under reduced pressure to give 4.63g of crude 8- (4-aminobutyl) -6, 7-dihydro-MeBmt]-1-cyclosporin [ formula: c₆₆H₁₂₂N₁₂O₁₂(ii) a Accurate quality: 1274.93, respectively; MS (M/z) 1275.71(M +1)⁺(ii) a HPLC RT: 11.95 min. (C8 reverse phase column: 250 mm; acetonitrile/water (0.05% trifluoroacetic acid); operating temperature: 64 ℃ C.; detector: 210nm)]。

[8- (4- (tert-Butoxycarbonyl) aminobutyl) -6, 7-dihydro-MeBmt ] -1-cyclosporin

Reacting [8- (4-aminobutyl) -6, 7-dihydro-MeBmt]-1-Cyclosporin (4.63g, 3.63mmol) was dissolved in tetrahydrofuran (50 ml). Saturated sodium bicarbonate solution (25ml) and di-tert-butyl dicarbonate (0.87g, 3.99mmol) were added. The mixture was stirred at room temperatureStirring for 2 hours. Most of the tetrahydrofuran was evaporated under reduced pressure. Ethyl acetate (30ml) and brine (30ml) were added and the mixture was separated. The organic layer was dried over magnesium sulfate and evaporated under reduced pressure. The residue was purified by chromatography (hexane/acetone) to give 3.00g of pure [8- (4- (tert-butoxycarbonyl) aminobutyl) -6, 7-dihydro-MeBmt]-1-cyclosporin [ formula: c₇₁H₁₃₀N₁₂O₁₄(ii) a Accurate quality: 1374.98, respectively; MS (M/z) 1375.64(M +1)⁺,1397.85(M+Na)⁺(ii) a HPLC RT: 17.81 min. (C8 reverse phase column: 250 mm; acetonitrile/water (0.05% trifluoroacetic acid); operating temperature: 64 ℃ C.; detector: 210nm)]。

[8- (4- (tert-Butoxycarbonyl) aminobutyl) -6, 7-dihydro-MeBmt ] -1- [ α -methylene-Sar ] -3-cyclosporin

N-butyllithium (2.65M,8.23ml, 21.82mmol) was added to a solution of diisopropylamine (3.09ml, 21.82mmol) in tetrahydrofuran (60ml) at-78 ℃ under nitrogen. After stirring the reaction mixture for 1 hour, [8- (4- (tert-butoxycarbonyl) aminobutyl) -6, 7-dihydro-MeBmt ] was added over 10 minutes]-1-Cyclosporin (3.00g, 2.18mmol) in tetrahydrofuran (20 ml). The mixture was stirred at-78 ℃ for 2 hours. After bubbling carbon dioxide gas into the reaction mixture for 30 minutes, the mixture was stirred at-78 ℃ for another 1 hour. The cooling bath was then removed and the reaction mixture was allowed to warm to room temperature to allow unreacted carbon dioxide to vent. The mixture was cooled to-78 ℃ and chloromethyl chloroformate (1.93ml, 21.82mmol) was added. The mixture was stirred and allowed to warm to room temperature overnight. Brine (10ml) was added to quench the reaction. Most of the tetrahydrofuran was removed under reduced pressure. Ethyl acetate (50ml) and brine (50ml) were added and the mixture was separated. The organic layer was dried over magnesium sulfate and evaporated under reduced pressure. The residue was purified by chromatography (hexane/acetone) to give 0.40g of pure [8- (4- (tert-butoxycarbonyl) aminobutyl) -6, 7-dihydro-MeBmt]-1- [ α -methylene-Sar]-3-cyclosporin [ formula: c₇₂H₁₃₀N₁₂O₁₄(ii) a Accurate quality: 1386.98, respectively; MS (M/z) 1387.61(M +1)⁺,1409.80(M+Na)⁺(ii) a HPLC RT: 19.01 min. (C8 reverse phase column: 250 mm; acetonitrile/water (0.05% trifluoroacetic acid); operating temperature: 64 ℃ C.; detector: 210nm)]。

[8- (4- (tert-Butoxycarbonyl) aminobutyl) -6, 7-dihydro-MeBmt ] -1- [ (S) - (4-hydroxybutylthio) methyl-Sar ] -3-cyclosporin

To [8- (4- (tert-butoxycarbonyl) aminobutyl) -6, 7-dihydro-MeBmt]-1- [ α -methylene-Sar]To a solution of (0.40g, 0.29mmol) of (E) -3-cyclosporin in methanol (15ml) were added (0.18ml, 1.73mmol) of 4-mercapto-1-butanol and (0.07g, 2.88mmol) of lithium hydroxide. The reaction mixture was stirred at room temperature for 7 hours. Most of the methanol was evaporated under reduced pressure. Ethyl acetate (50ml) and brine (50ml) were added and the mixture was separated. The organic layer was dried over magnesium sulfate and evaporated under reduced pressure. The residue was purified by chromatography (hexane/acetone from 90/10 to 75/25) to yield 200mg of pure [8- (4- (tert-butoxycarbonyl) aminobutyl) -6, 7-dihydro-MeBmt]-1- [ (S) - (4-hydroxybutylthio) methyl-Sar]-3-cyclosporin [ formula: c₇₆H₁₄₀N₁₂O₁₅S; accurate quality: 1493.03, respectively; MS (M/z) 1493.63(M +1)⁺,1515.88(M+Na)⁺(ii) a HPLC RT: 17.65 min. (C8 reverse phase column: 250 mm; acetonitrile/water (0.05% trifluoroacetic acid); operating temperature: 64 ℃ C.; detector: 210nm)]。

[8- (4-Acylaminobutyl) -6, 7-dihydro-MeBmt ] -1- [ (S) - (4-hydroxybutylthio) methyl-Sar ] -3-cyclosporin

Reacting [8- (4- (tert-butoxycarbonyl) aminobutyl) -6, 7-dihydro-MeBmt]-1- [ (S) - (4-hydroxybutylthio) methyl-Sar]-3-Cyclosporin (0.20g, 0.16mmol) solutionDissolved in dichloromethane (15ml) and placed in an ice-water bath. Trifluoroacetic acid (5ml) was added. The mixture was stirred at 0 ℃ for 1 hour. Another portion of dichloromethane (20ml) was added. The mixture was washed with brine (30ml), saturated sodium bicarbonate solution (30ml), dried over magnesium sulfate and evaporated under reduced pressure to give crude [8- (4-aminobutyl) -6, 7-dihydro-MeBmt]-1- [ (S) - (4-hydroxybutylthio) methyl-Sar]-3-cyclosporin [ formula: c₇₁H₃₂N₁₂O₁₃S; accurate quality: 1392.98, respectively; MS (M/z) 1393.80(M +1)⁺(ii) a HPLCRT: 11.38 min. (C8 reverse phase column: 250 mm; acetonitrile/water (0.05% trifluoroacetic acid); operating temperature: 64 ℃ C.; detector: 210nm)]. Crude [8- (4-aminobutyl) -6, 7-dihydro-MeBmt]-1- [ (S) - (4-hydroxybutylthio) methyl-Sar]-3-Cyclosporin was dissolved in dichloromethane (15 ml). Acetic acid (48mg, 0.80mmol), HBTU (0.18g, 0.48mmol), 1-hydroxybenzotriazole (0.06g, 0.48mmol) and pyridine (0.50ml) were added. The mixture was stirred at room temperature for 2 hours. Dichloromethane (30ml) and brine (50ml) were then added and separated. The dichloromethane layer was dried over magnesium sulfate and evaporated under reduced pressure. The residue was purified by chromatography (dichloromethane/methanol) to give 27mg of [8- (4-acetamidobutyl) -6, 7-dihydro-MeBmt]-1- [ (S) - (4-hydroxybutylthio) methyl-Sar]-3-cyclosporin [ formula: c₇₃H₁₃₄N₁₂O₁₄S; accurate quality: 1434.99, respectively; MS (M/z) 1435.69(M +1)⁺,1457.87(M+Na)⁺(ii) a HPLC RT: 16.01 min. (C8 reverse phase column: 250 mm; acetonitrile/water (0.05% trifluoroacetic acid); operating temperature: 64 ℃ C.; detector: 210nm)]。

Example 36

[8- (2-Acetylaminoethyl) -6, 7-dihydro-MeBmt]-1- [ (S) - (4-hydroxybutylthio) methyl-Sar]-3- [ (gamma-hydroxy) -N-MeLeu]-4-Cyclosporin

[ 3-chloroacetyl-MeBmt ] -1- [ (gamma-hydroxy) -N-MeLeu ] -4-cyclosporin

At-35 deg.C, to [ (gamma-hydroxy) -N-MeLeu]To a solution of (12.17g, 1.00mmol) of (E) -4-cyclosporin in N, N-dimethylformamide (300ml) were added N, N-dimethylaminopyridine (0.12g, 0.10mmol), anhydrous pyridine (16.00g, 0.20mol) and chloroacetic anhydride (72.05g, 0.54 mol). The reaction mixture was stirred at room temperature overnight. The mixture was poured into 800ml of ice water and stirred until the ice melted. Ethyl acetate (500ml) was added and the mixture was separated. The ethyl acetate layer was washed with water (100ml), saturated sodium bicarbonate solution (100ml), brine (100ml), dried over magnesium sulfate and evaporated under reduced pressure. The residue was purified by chromatography (hexane/acetone) to give 9.40g of pure [ 3-chloroacetyl-MeBmt]-1- [ (γ -hydroxy) -N-MeLeu]-4-cyclosporin [ formula: c₆₄H₁₁₃ClN₁₁O₁₄(ii) a Accurate quality: 1294.82, respectively; MS (M/z) 1295.50(M +1)⁺]。

[ (gamma-hydroxy) -N-MeLeu ] -4-cyclosporin is prepared by Sebekia benihana Biotransformation according to the method described by Kuhnt M. et al, 1996, Microbial Biotransformation Products of Cyclosporin A, J.antibiotics,49(8), 781.

[ (3R,4R) -3-Chloroacetoxy-4-methyl-6-oxo-N-Menle ] -1- [ (γ -hydroxy) -N-MeLeu ] -4-Cyclosporin

To [ 3-chloroacetyl-MeBmt]-1- [ (γ -hydroxy) -N-MeLeu]To a solution of-4-cyclosporin (9.50g, 7.34mmol) in dioxane (125ml) was added water (100ml), osmium (VIII) oxide solution (0.4% in water, 35ml) and sodium metaperiodate (6.60g, 30.90 mmol). The reaction mixture was stirred at room temperature for 5 hours. Ethyl acetate (100ml) and brine (100ml) were added and the mixture was separated. The organic layer was washed with saturated sodium bicarbonate solution, brine, dried over magnesium sulfate and evaporated under reduced pressure. The residue was purified by chromatography (hexane/acetone) to give 5.00g of pure [ (3R,4R) -3-chloroacetoxy-4-methylradical-6-oxo-N-Menle]-1- [ (γ -hydroxy) -N-MeLeu]-4-cyclosporin [ formula: c₆₂H₁₀₉ClN₁₁O₁₅(ii) a Accurate quality: 1282.78, respectively; MS (M/z) 1283.47(M +1)⁺(ii) a HPLC RT: 14.74 min. (C8 reverse phase column: 250 mm; acetonitrile/water (0.05% trifluoroacetic acid); operating temperature: 64 ℃ C.; detector: 210nm)]。

[ 3-Chloroacyl-8-cyanomethyl-MeBmt ] -1- [ (Gamma-hydroxy) -N-MeLeu ] -4-Cyclosporin

To a solution of (3-cyanopropyl) triphenyl phosphonium bromide (3.07g, 7.48mmol) in anhydrous tetrahydrofuran (120ml) was added sodium bis (trimethylsilyl) amide (1.00M in tetrahydrofuran, 14ml, 14.00mmol) under nitrogen. The reaction mixture was stirred at room temperature for one hour and cooled to-30 ℃. Adding [ (3R,4R) -3-chloroacetoxy-4-methyl-6-oxo-N-Menle]-1- [ (γ -hydroxy) -N-MeLeu]-4-Cyclosporin (4.80g, 3.74mmol) in dry tetrahydrofuran (15 ml). The mixture was stirred at-30 ℃ for a further 2 hours. Saturated ammonium chloride solution (20ml) was then added to quench the reaction. Most of the tetrahydrofuran was evaporated under reduced pressure. Ethyl acetate (100ml) and brine (100ml) were added and the mixture was separated. The organic layer was dried over magnesium sulfate and evaporated under reduced pressure. The residue was purified by chromatography (dichloromethane/methanol) to give 2.00g of pure [ 3-chloroacetyl-8-cyanomethyl-MeBmt]-1- [ (γ -hydroxy) -N-MeLeu]-4-cyclosporin [ formula: c₆₆H₁₁₄ClN₁₂O₁₄(ii) a Accurate quality: 1333.83, respectively; MS (M/z) 1334.56(M +1)⁺]。

[ 3-acetyl-8- (2-aminoethyl) -6, 7-dihydro-MeBmt ] -1- [ (Gamma-hydroxy) -N-MeLeu ] -4-Cyclosporin

To [ 3-chloroacetyl group-8-cyanomethyl-MeBmt]-1- [ (γ -hydroxy) -N-MeLeu]To a solution of (0.85g, 0.63mmol) of (4-cyclosporin in) in methanol (100ml) was added nickel (II) chloride hexahydrate (0.19g, 0.81 mmol). Sodium borohydride (0.38g, 10.00mmol) was then added portionwise over 30 minutes. After stirring the mixture at room temperature for a further 1 hour, most of the methanol was evaporated under reduced pressure. Ethyl acetate (50ml) and saturated sodium bicarbonate solution (50ml) were added to the solution, and the mixture was separated. The organic layer was washed with brine, dried over magnesium sulfate and evaporated under reduced pressure. The residue was purified by chromatography (dichloromethane/methanol) to give 0.61g of [ 3-acetyl-8- (2-aminoethyl) -6, 7-dihydro-MeBmt]-1- [ (γ -hydroxy) -N-MeLeu]-4-cyclosporin [ formula: c₆₆H₁₂₀N₁₂O₁₄(ii) a Accurate quality: 1304.90, respectively; MS (M/z) 1305.68(M +1)⁺(ii) a HPLC RT: 12.80 min. (C8 reverse phase column: 250 mm; acetonitrile/water (0.05% trifluoroacetic acid); operating temperature: 64 ℃ C.; detector: 210nm)]。

[8- (2-Acetylaminoethyl) -3-acetyl-6, 7-dihydro-MeBmt ] -1- [ (Gamma-hydroxy) -N-MeLeu ] -4-Cyclosporin

To [ 3-acetyl-8- (2-aminoethyl) -6, 7-dihydro-MeBmt]-1- [ (γ -hydroxy) -N-MeLeu]To a solution of (0.80g, 0.61mmol) of (4-cyclosporin and (0.11g, 1.83mmol) of acetic acid in dichloromethane (60ml) were added diisopropylethylamine (0.25g, 1.93mmol) and HATU (0.70g, 1.83 mmol). The reaction mixture was stirred at room temperature for 3 hours, then washed with saturated sodium bicarbonate solution, brine, dried over magnesium sulfate and evaporated under reduced pressure. The residue was purified by chromatography (dichloromethane/methanol) to give 0.55g of pure [8- (2-acetamidomethyl) -3-acetyl-6, 7-dihydro-MeBmt]-1- [ (γ -hydroxy) -N-MeLeu]-4-cyclosporin [ formula: c₆₈H₁₂₂N₁₂O₁₅(ii) a Accurate quality: 1346.92, respectively; MS (M/z) 1347.63(M +1)⁺(ii) a HPLC RT:13.74 min. (C8 reverse phase column: 250 mm; acetonitrile/water (0.05% trifluoroacetic acid); operating temperature: 64 ℃ C.; detector: 210nm)]。

[8- (2-Acetylaminoethyl) -6, 7-dihydro-MeBmt ] -1- [ (Gamma-hydroxy) -N-MeLeu ] -4-Cyclosporin

Reacting [8- (2-acetamido ethyl) -3-acetyl-6, 7-dihydro-MeBmt]-1- [ (γ -hydroxy) -N-MeLeu]-4-Cyclosporin (0.50g, 0.37mmol) was dissolved in methanol (50 ml). Tetramethylammonium hydroxide pentahydrate (0.22g, 1.21mmol) was added. The mixture was stirred at room temperature for two days. Most of the methanol is then evaporated. Ethyl acetate (100ml) and brine (100ml) were added and the mixture was separated. The organic layer was dried over magnesium sulfate and evaporated under reduced pressure. The residue was purified by chromatography (dichloromethane/methanol) to give 0.22g of [8- (2-acetamidomethyl) -6, 7-dihydro-MeBmt]-1- [ (γ -hydroxy) -N-MeLeu]-4-cyclosporin [ formula: c₆₆H₁₂₀N₁₂O₁₄(ii) a Accurate quality: 1304.90, respectively; MS (M/z) 1305.72(M +1)⁺(ii) a HPLC RT: 12.80 min. (C8 reverse phase column: 250 mm; acetonitrile/water (0.05% trifluoroacetic acid); operating temperature: 64 ℃ C.; detector: 210nm)]。

[8- (2-Acetylaminoethyl) -6, 7-dihydro-MeBmt ] -1- [ α -methylene-Sar ] -3- [ (gamma-hydroxy) -N-MeLeu ] -4-Cyclosporin

N-butyllithium (2.65M, 5.3ml, 14.04mmol) was added to a solution of diisopropylamine (2.11ml, 14.85mmol) in tetrahydrofuran (50ml) at-78 ℃ under nitrogen. After stirring the reaction mixture for 1 hour, [8- (2-acetamidomethyl) -6, 7-dihydro-MeBmt ] was added over 10 minutes]-1- [ (γ -hydroxy) -N-MeLeu]-4-Cyclosporin (1.60g, 1.23mmol) in tetrahydrofuran (6 ml). The mixture was stirred at-78 ℃ for three hours. After bubbling carbon dioxide gas into the reaction mixture for 30 minutes, the mixture was stirred at-78 ℃ for another 1 hour. Then move backwardsThe reaction mixture was allowed to warm to room temperature to allow unreacted carbon dioxide to vent off the cooling bath. The mixture was cooled to-78 ℃ and chloromethyl chloroformate (1.32ml, 14.85mmol) was added. The mixture was stirred and allowed to warm to room temperature overnight. Brine (5ml) was added to quench the reaction. Most of the tetrahydrofuran was removed under reduced pressure. Ethyl acetate (50ml) and brine (50ml) were added and the mixture was separated. The organic layer was dried over magnesium sulfate and evaporated under reduced pressure. The residue was purified by chromatography (hexane/acetone) to give 0.50g of pure [8- (2-acetamidomethyl) -6, 7-dihydro-MeBmt]-1- [ α -methylene-Sar]-3- [ (γ -hydroxy) -N-MeLeu]-4-cyclosporin [ formula: c₆₇H₁₂₀N₁₂O₁₄(ii) a Accurate quality: 1316.90, respectively; MS (M/z) 1317.66(M +1)⁺(ii) a HPLC RT: 14.32 min. (C8 reverse phase column: 250 mm; acetonitrile/water (0.05% trifluoroacetic acid); operating temperature: 64 ℃ C.; detector: 210nm)]。

[8- (2-Acetylaminoethyl) -6, 7-dihydro-MeBmt ] -1- [ (S) - (4-hydroxybutylthio) methyl-Sar ] -3- [ (gamma-hydroxy) -N-MeLeu ] -4-Cyclosporin

To [8- (2-acetamidoethyl) -6, 7-dihydro-MeBmt]-1- [ α -methylene-Sar]-3- [ (γ -hydroxy) -N-MeLeu]To a solution of (0.30g, 0.23mmol) of (E) -4-cyclosporin in methanol (20ml) were added (0.14ml, 1.38mmol) of 4-mercapto-1-butanol and (54mg, 2.31mmol) of lithium hydroxide. The reaction mixture was stirred at room temperature for 5 hours. Most of the methanol was evaporated under reduced pressure. Ethyl acetate (30ml) and brine (30ml) were added and the mixture was separated. The organic layer was dried over magnesium sulfate and evaporated under reduced pressure. The residue was purified by chromatography (dichloromethane/methanol) to give 120mg of pure [8- (2-acetamidomethyl) -6, 7-dihydro-MeBmt]-1- [ (S) - (4-hydroxybutylthio) methyl-Sar]-3- [ (γ -hydroxy) -N-MeLeu]-4-cyclosporin [ formula: c₇₁H₁₃₀N₁₂O₁₅S; accurate quality: 1422.95, respectively; MS (M/z) 1423.74(M +1)⁺(ii) a HPLC RT: 11.98 min. (C8 reverse phase column: 250 mm; acetonitrile/water (0.05% trifluoroacetic acid)) (ii) a Operating temperature: 64 ℃; a detector: 210nm)]。

Example 37

[8- (3-Acetylaminopropyl) -6, 7-dihydro-MeBmt]-1- [ (R) - (3-hydroxypropyl) thio-Sar]-3-Cyclosporium sp Bacterins

p-Toluenesulfothiosulfonic acid potassium salt (4.5g, 19.91mmol) and 3-bromo-1-propanol (2.8g, 20.29mmol) were added to ethanol (50 ml). The reaction mixture was stirred and heated to reflux for 4 hours. Most of the ethanol was evaporated under reduced pressure. The residue was mixed with ethyl acetate (100 ml). The ethyl acetate layer was washed with brine, dried over magnesium sulfate and evaporated to give 14.79g of crude S- (3-hydroxypropyl) 4-methylthiobenzenesulfonate.

S- (3- ((tetrahydro-2H-pyran-2-yl) oxy) propyl) 4-methylthiobenzenesulfonate

S- (3-hydroxypropyl) 4-methylthiobenzenesulfonate (7.20g, 29.26mmol) was dissolved in dichloromethane (100 ml). 3, 4-dihydro-2H-pyran (3.00g, 35.66mmol) and p-toluenesulfonic acid monohydrate (1.00g, 5.26mmol) were added. The mixture was stirred at room temperature overnight. The dichloromethane was washed with sodium bicarbonate solution and brine, dried over magnesium sulfate and evaporated under reduced pressure. The residue was purified by chromatography to give 6.50g of pure S- (3- ((tetrahydro-2H-pyran-2-yl) oxy) propyl) 4-methylthiobenzenesulfonate. [ molecular formula: c₁₅H₂₂O₄S₂(ii) a Accurate quality: 330.10, respectively; MS (M/z) 330.95(M +1)⁺.

[8- (3-aminopropyl) -6, 7-dihydro-MeBmt ] -1- [ (R) - (3- ((tetrahydro-2H-pyran-2-yl) oxy) propyl) thio-Sar ] -3-cyclosporin

N-butyllithium (2.60M, 6.40ml, 16.64mmol) was added to a solution of diisopropylamine (2.32ml, 16.33mmol) in tetrahydrofuran (50ml) at-78 ℃ under nitrogen. After stirring the reaction mixture for 1 hour, [8- (3- (aminopropyl) -6, 7-dihydro-MeBmt ] was added over 10 minutes]-1-Cyclosporin (1.50g, 1.11mmol) in tetrahydrofuran (15 ml). The mixture was stirred at-78 ℃ for 2 hours. A solution of S- (3- ((tetrahydro-2H-pyran-2-yl) oxy) propyl) 4-methylthiobenzenesulfonate (2.70g, 8.18mmol) in tetrahydrofuran (10ml) was added and the mixture was stirred at-78 ℃ for a further 2 hours. The cooling bath was then removed and the reaction mixture was allowed to warm to room temperature and stirred for an additional 2 hours. Saturated ammonium chloride solution (20ml) was added to quench the reaction. Most of the tetrahydrofuran was removed under reduced pressure. Ethyl acetate (50ml) and brine (50ml) were added and the mixture was separated. The organic layer was dried over magnesium sulfate and evaporated under reduced pressure. The residue was purified by chromatography (dichloromethane/methanol) to give 0.38g of [8- (3-aminopropyl) -6, 7-dihydro-MeBmt]-1- [ (R) - (3- ((tetrahydro-2H-pyran-2-yl) oxy) propyl) thio-Sar]-3-cyclosporin [ formula: c₇₃H₁₃₄N₁₂O₁₄S; accurate quality: 1434.99, respectively; MS (M/z) 1435.70(M +1)⁺(ii) a HPLC RT:14.93 min. (C8 reverse phase column: 250 mm; acetonitrile/water (0.05% trifluoroacetic acid); operating temperature: 64 ℃ C.; detector: 210nm)]。

[8- (3-Acetylaminopropyl) -6, 7-dihydro-MeBmt ] -1- [ (R) - (3- ((tetrahydro-2H-pyran-2-yl) oxo) propyl) thio-Sar ] -3-cyclosporin

To [8- (3-aminopropyl) -6, 7-dihydro-MeBmt]-1- [ (R) - (3- ((tetrahydro-2H-pyran-2-yl) oxy) propyl) thio-Sar]-3-Cyclosporin (0.50g, 0.35mmol) and acetic acidTo a solution of (50mg, 0.83mmol) in dichloromethane (50ml) were added diisopropylethylamine (90mg, 0.70mmol), HOBT (85mg, 0.56mmol) and HATU (237mg, 0.63 mmol). The reaction mixture was stirred at room temperature for 2 hours, then washed with saturated sodium bicarbonate solution, brine, dried over magnesium sulfate and evaporated under reduced pressure. The residue was purified by chromatography (dichloromethane/methanol) to give 142mg of pure [8- (3-acetamidopropyl) -6, 7-dihydro-MeBmt]-1- [ (R) - (3- ((tetrahydro-2H-pyran-2-yl) oxy) propyl) thio-Sar]-3-cyclosporin [ formula: c₇₅H₁₃₅N₁₂O₁₅S; accurate quality: 1477.00, respectively; MS (M/z) 1477.65(M +1)⁺. HPLC RT 18.17min (C8 reverse phase column: 250 mm; acetonitrile/water (0.05% trifluoroacetic acid); operating temperature: 64 ℃ C.; detector: 210nm)]。

[8- (3-Acetylaminopropyl) -6, 7-dihydro-MeBmt ] -1- [ (R) - (3-hydroxypropyl) thio-Sar ] -3-cyclosporin

To [8- (3-acetamidopropyl) -6, 7-dihydro-MeBmt]-1- [ (R) - (3- ((tetrahydro-2H-pyran-2-yl) oxo) propyl) thio-Sar]To a solution of-3-cyclosporin (0.14g, 0.35mmol) in methanol (15ml) was added Dowex-50WX4(200 mg). The reaction mixture was stirred at room temperature for 4 hours, and then filtered. Most of the methanol was evaporated under reduced pressure and the residue was purified by chromatography (dichloromethane/methanol) to give 48mg of pure [8- (3-acetamidopropyl) -6, 7-dihydro-MeBmt]-1- [ (R) - (3-hydroxypropyl) thio-Sar]-3-cyclosporin [ formula: c₇₀H₁₂₈N₁₂O₁₄S; accurate quality: 1392.94, respectively; MS (M/z) 1393.66(M +1)⁺. HPLC RT:15.38min (C8 reverse phase column: 250 mm; acetonitrile/water (0.05% trifluoroacetic acid); operating temperature: 64 ℃ C.; detector: 210nm)]。

Example 36

[8- (2-Acetylaminoethyl) -6, 7-dihydro-MeBmt]-1-[ (R) - (4-hydroxybutyl) thio-Sar]-3-Cyclosporium sp Bacterins

4- (tosylthio) butyl benzoate

p-Toluenesulfonic acid potassium salt (20.00g, 88.36mmol) and 4-chloro-1-butanol (9.60g, 88.36mmol) were added to ethanol (160 ml). The reaction mixture was stirred and heated to reflux for 4 hours. Most of the ethanol was evaporated under reduced pressure. The residue was mixed with ethyl acetate (100 ml). The ethyl acetate layer was washed with brine, dried over magnesium sulfate and evaporated to give 14.79g of crude S- (4-hydroxybutyl) 4-methylthiobenzenesulfonate.

S- (4-hydroxybutyl) 4-methylthiobenzenesulfonate (14.79g, 56.90mmol) was dissolved in acetone (60 ml). Benzoic anhydride (25.74, 113.80mmol), 4- (dimethylamino) pyridine (1.39g, 11.38mmol) and pyridine (30ml) were added. The mixture was stirred at room temperature overnight. Most of the acetone and pyridine were evaporated under reduced pressure. Ethyl acetate (100ml) was added. The ethyl acetate layer was washed with a hydrochloric acid solution (1.00N), brine, saturated sodium bicarbonate solution and brine, dried over magnesium sulfate and evaporated under reduced pressure. The residue was purified by chromatography (hexane/acetone) to give 3.30g of pure 4- (tosylthio) butyl benzoate [ formula: c₁₈H₂₀O₄S₂(ii) a Accurate quality: 364.08, respectively; MS (M/z) 364.57(M +1)⁺(ii) a HPLC RT: 19.44 min. (C8 reverse phase column: 250 mm; acetonitrile/water (0.05% trifluoroacetic acid); operating temperature: 64 ℃ C.; detector: 210nm)]。

[8- (2- (tert-Butoxycarbonyl) aminoethyl) -6, 7-dihydro-MeBmt]-1- [ (R) - (4- (benzoyloxy) butan Yl) thio-Sar]-3-Cyclosporin

At-78 deg.C,N-butyllithium (2.60M, 6.28ml, 16.33mmol) was added to a solution of diisopropylamine (2.32ml, 16.33mmol) in tetrahydrofuran (50ml) under nitrogen. After stirring the reaction mixture for 1 hour, [8- (2- (tert-butoxycarbonyl) aminoethyl) -6, 7-dihydro-MeBmt was added over 10 minutes]-1-Cyclosporin (2.20g, 1.63mmol) in tetrahydrofuran (15 ml). The mixture was stirred at-78 ℃ for 2 hours. A solution of 4- (tosylthio) butyl benzoate (3.56g, 9.78mmol) in tetrahydrofuran (10ml) was added and the mixture was stirred at-78 ℃ for a further 2 h. The cooling bath was then removed and the reaction mixture was allowed to warm to room temperature and stirred for an additional 2 hours. Saturated ammonium chloride solution (20ml) was added to quench the reaction. Most of the tetrahydrofuran was removed under reduced pressure. Ethyl acetate (50ml) and brine (50ml) were added and the mixture was separated. The organic layer was dried over magnesium sulfate and evaporated under reduced pressure to give 0.38g of crude [8- (2- (tert-butoxycarbonyl) aminoethyl) -6, 7-dihydro-MeBmt]-1- [ (R) - (4- (benzoyloxy) butyl) thio-Sar]-3-cyclosporin [ formula: c₈₀H₁₃₈N₁₂O₁₆S; accurate quality: 1555.01, respectively; MS (M/z) 1555.72(M +1)⁺(ii) a HPLC RT: 19.21 min. (C8 reverse phase column: 250 mm; acetonitrile/water (0.05% trifluoroacetic acid); operating temperature: 64 ℃ C.; detector: 210nm)]。

[8- (2- (tert-Butoxycarbonyl) aminoethyl) -6, 7-dihydro-MeBmt ] -1- [ (R) - (4-hydroxybutyl) thio-Sar ] -3-cyclosporin

[8- (2- (tert-Butoxycarbonyl) aminoethyl) -6, 7-dihydro-MeBmt from the previous step]-1- [ (R) - (4- (benzoyloxy) butyl) thio-Sar]-3-Cyclosporin was dissolved in methanol (10 ml). Lithium hydroxide (2.00g, 83.33mmol) and water (10ml) were added. The mixture was stirred at room temperature for 2 hours. Most of the methanol was then evaporated under reduced pressure. Ethyl acetate (50ml) and brine (50ml) were added and separated. The ethyl acetate layer was dried over magnesium sulfate and evaporated under reduced pressure. The residue was purified by chromatography (hexane/acetone) to give 0.38g of pure [8- (2- (tert-butoxycarbonyl) aminoethylether)Yl) -6, 7-dihydro-MeBmt]-1- [ (R) - (4-hydroxybutyl) thio-Sar]-3-cyclosporin [ formula: c₇₃H₁₃₄N₁₂O₁₅S; accurate quality: 1450.98, respectively; MS (M/z) 1451.68(M +1)⁺,1473.79(M+Na)⁺(ii) a HPLC RT: 17.06 min. (C8 reverse phase column: 250 mm; acetonitrile/water (0.05% trifluoroacetic acid); operating temperature: 64 ℃ C.; detector: 210nm)]。

[8- (2-aminoethyl) -6, 7-dihydro-MeBmt ] -1- [ (R) - (4-hydroxybutyl) thio-Sar ] -3-cyclosporin

Reacting [8- (2- (tert-butoxycarbonyl) aminoethyl) -6, 7-dihydro-MeBmt]-1- [ (R) - (4-hydroxybutyl) thio-Sar]-3-Cyclosporin (0.38g, 0.26mmol) was dissolved in dichloromethane (12ml) and placed in an ice-water bath. Trifluoroacetic acid (4ml) was added. The mixture was stirred at 0 ℃ for three hours. Another portion of dichloromethane (50ml) was added. The mixture was washed with brine (50ml), saturated sodium bicarbonate solution (50ml), dried over magnesium sulfate and evaporated under reduced pressure to give crude [8- (2-aminoethyl) -6, 7-dihydro-MeBmt]-1- [ (R) - (4-hydroxybutyl) thio-Sar]-3-cyclosporin [ formula: c₆₈H₁₂₆N₁₂O₁₃S; accurate quality: 1350.93, respectively; MS (M/z) 1351.67(M +1)⁺(ii) a HPLC RT: 10.72 min. (C8 reverse phase column: 250 mm; acetonitrile/water (0.05% trifluoroacetic acid); operating temperature: 64 ℃ C.; detector: 210nm)]。

[8- (2-Acetylaminoethyl) -6, 7-dihydro-MeBmt ] -1- [ (R) - (4-hydroxybutyl) thio-Sar ] -3-Cyclosporin

Crude [8- (2-aminoethyl) -6, 7-dihydro-MeBmt]-1- [ (R) - (4-hydroxybutyl) thio-Sar]-3-Cyclosporin was dissolved in dichloromethane (15 ml). Acetic acid (0.08g, 1.33mmol), HBTU (0.30g, 0.79mmol), and 1-hydroxy group were addedBenzotriazole (0.11g, 0.79mmol) and diisopropylethylamine (0.50 ml). The mixture was stirred at room temperature for 2 hours. Most of the solvent was then evaporated under reduced pressure. Ethyl acetate (50ml) and brine (50ml) were added and separated. The ethyl acetate layer was dried over magnesium sulfate and evaporated under reduced pressure. The residue was purified by chromatography (hexane/acetone) to give 25mg of pure [8- (2-acetamidomethyl) -6, 7-dihydro-MeBmt]-1- [ (R) - (4-hydroxybutyl) thio-Sar]-3-cyclosporin [ formula: c₇₀H₁₂₉N₁₂O₁₄S; accurate quality: 1392.94, respectively; MS (M/z) 1393.98(M +1)⁺,1415.92(M+Na)⁺(ii) a HPLC RT: 15.09 min. (C8 reverse phase column: 250 mm; acetonitrile/water (0.05% trifluoroacetic acid); operating temperature: 64 ℃ C.; detector: 210nm)]。

Example 39

[8- (2-Acetylaminoethyl) -6, 7-dihydro-MeBmt]-1- [ (R) - (5-hydroxypentyl) thio-Sar]-3-Cyclosporium sp Bacterins

5- (tosylthio) pentylbenzoate

p-Toluenesulfonic acid potassium salt (11.80g, 47.71mmol) and 5-chloro-1-pentanol (5.84g, 47.71mmol) were added to ethanol (100 ml). The reaction mixture was stirred and heated to reflux overnight. Most of the ethanol was evaporated under reduced pressure. The residue was mixed with ethyl acetate (100 ml). The ethyl acetate layer was washed with brine, dried over magnesium sulfate and evaporated to give 9.16g of crude S- (5-hydroxypentyl) 4-methylthiobenzenesulfonate.

S- (5-hydroxypentyl) 4-methylthiobenzenesulfonate (9.16g, 33.42mmol) was dissolved in acetone (60 ml). Benzoic anhydride (15.12, 66.83mmol), 4- (dimethylamino) pyridine (0.82g, 6.71mmol) and pyridine (30ml) were added. The mixture was stirred at room temperature overnight. Most of the acetone and pyridine were evaporated under reduced pressure. Ethyl acetate (100ml) was added. With hydrochloric acidThe ethyl acetate layer was washed with brine, dried over magnesium sulfate and evaporated under reduced pressure. The residue was purified by chromatography (hexane/acetone) to give 7.78g of pure 5- (tosylthio) pentylbenzoate [ formula: c₁₉H₂₂O₄S₂(ii) a Accurate quality: 378.10, respectively; MS (M/z) 378.57(M +1)⁺(ii) a HPLC RT: 20.24 min. (C8 reverse phase column: 250 mm; acetonitrile/water (0.05% trifluoroacetic acid); operating temperature: 64 ℃ C.; detector: 210nm)]。

[8- (2- (tert-Butoxycarbonyl) aminoethyl) -6, 7-dihydro-MeBmt ] -1- [ (R) - (5- (benzoyloxy) pentyl) thio-Sar ] -3-cyclosporin

N-butyllithium (2.65M, 6.16ml, 16.33mmol) was added to a solution of diisopropylamine (2.32ml, 16.33mmol) in tetrahydrofuran (50ml) at-78 ℃ under nitrogen. After stirring the reaction mixture for 1 hour, [8- (2- (tert-butoxycarbonyl) aminoethyl) -6, 7-dihydro-MeBmt was added over 10 minutes]-1-Cyclosporin (2.20g, 1.63mmol) in tetrahydrofuran (15 ml). The mixture was stirred at-78 ℃ for 2 hours. A solution of 5- (tosylthio) pentylbenzoate (3.71g, 9.80mmol) in tetrahydrofuran (10ml) was added and the mixture was stirred at-78 ℃ for an additional 2 hours. The cooling bath was then removed and the reaction mixture was allowed to warm to room temperature and stirred for an additional 2 hours. Saturated ammonium chloride solution (20ml) was added to quench the reaction. Most of the tetrahydrofuran was removed under reduced pressure. Ethyl acetate (80ml) and brine (80ml) were added to the solution, and the mixture was separated. The organic layer was dried over magnesium sulfate and evaporated under reduced pressure to give 0.38g of crude [8- (2- (tert-butoxycarbonyl) aminoethyl) -6, 7-dihydro-MeBmt]-1- [ (R) - (5- (benzoyloxy) pentyl) thio-Sar]-3-cyclosporin [ formula: c₈₁H₁₄₀N₁₂O₁₆S; accurate quality: 1569.02, respectively; MS (M/z) 1569.73(M +1)⁺(ii) a HPLC RT: 19.56 min. (C8 reverse phase column: 250 mm; acetonitrile/water (0.05% trifluoroacetic acid); operating temperature: 64 ℃ C.; detector: 21;)0nm)]。

[8- (2- (tert-Butoxycarbonyl) aminoethyl) -6, 7-dihydro-MeBmt ] -1- [ (R) - (5-hydroxypentyl) thio-Sar ] -3-cyclosporin

[8- (2- (tert-Butoxycarbonyl) aminoethyl) -6, 7-dihydro-MeBmt from the previous step]-1- [ (R) - (5- (benzoyloxy) pentyl) thio-Sar]-3-Cyclosporin was dissolved in methanol (25 ml). Lithium hydroxide (1.25g, 52.08mmol) and water (25ml) were added. The mixture was stirred at room temperature for 2 hours. Most of the methanol was then evaporated under reduced pressure. Ethyl acetate (50ml) and brine (50ml) were added and separated. The ethyl acetate layer was dried over magnesium sulfate and evaporated under reduced pressure. The residue was purified by chromatography (hexane/acetone) to give 0.22g of pure [8- (2- (tert-butoxycarbonyl) aminoethyl) -6, 7-dihydro-MeBmt]-1- [ (R) - (5-hydroxypentyl) thio-Sar]-3-cyclosporin [ formula: c₇₄H₁₃₆N₁₂O₁₅S; accurate quality: 1465.00, respectively; MS (M/z) 1465.68(M +1)⁺,1487.79(M+Na)⁺(ii) a HPLC RT: 17.44 min. (C8 reverse phase column: 250 mm; acetonitrile/water (0.05% trifluoroacetic acid); operating temperature: 64 ℃ C.; detector: 210nm)]。

[8- (2-aminoethyl) -6, 7-dihydro-MeBmt ] -1- [ (R) - (5-hydroxypentyl) thio-Sar ] -3-Cyclosporin

Reacting [8- (2- (tert-butoxycarbonyl) aminoethyl) -6, 7-dihydro-MeBmt]-1- [ (R) - (5-hydroxypentyl) thio-Sar]-3-Cyclosporin (0.22g, 0.15mmol) was dissolved in dichloromethane (6ml) and placed in an ice-water bath. Trifluoroacetic acid (2ml) was added. The mixture was stirred at 0 ℃ for three hours. Another portion of dichloromethane (50ml) was added. The dichloromethane layer was washed with brine (50ml), saturated sodium bicarbonate solution (50ml), dried over magnesium sulfate and evaporated under reduced pressure to give crude [8-, (50ml)2-aminoethyl) -6, 7-dihydro-MeBmt]-1- [ (R) - (5-hydroxypentyl) thio-Sar]-3-cyclosporin [ formula: c₆₉H₁₂₈N₁₂O₁₃S; accurate quality: 1364.94, respectively; MS (M/z) 1365.67(M +1)⁺(ii) a HPLC RT: 11.53 min. (C8 reverse phase column: 250 mm; acetonitrile/water (0.05% trifluoroacetic acid); operating temperature: 64 ℃ C.; detector: 210nm)]。

[8- (2-Acetylaminoethyl) -6, 7-dihydro-MeBmt ] -1- [ (R) - (5-hydroxypentyl) thio-Sar ] -3-Cyclosporin

Crude [8- (2-aminoethyl) -6, 7-dihydro-MeBmt]-1- [ (R) - (5-hydroxypentyl) thio-Sar]-3-Cyclosporin was dissolved in dichloromethane (15 ml). Acetic acid (0.05g, 0.83mmol), HBTU (0.17g, 0.45mmol), 1-hydroxybenzotriazole (0.06g, 0.45mmol) and pyridine (0.50ml) were added. The mixture was stirred at room temperature for three hours. Most of the solvent was then evaporated under reduced pressure. Ethyl acetate (50ml) and brine (50ml) were added and separated. The ethyl acetate layer was dried over magnesium sulfate and evaporated under reduced pressure. The residue was purified by chromatography (hexane/acetone) to give 12mg of pure [8- (2-acetamidomethyl) -6, 7-dihydro-MeBmt]-1- [ (R) - (5-hydroxypentyl) thio-Sar]-3-cyclosporin [ formula: c₇₁H₁₃₀N₁₂O₁₄S; accurate quality: 1406.96, respectively; MS (M/z) 1407.77(M +1)⁺,1429.96(M+Na)⁺(ii) a HPLC RT: 15.72 min. (C8 reverse phase column: 250 mm; acetonitrile/water (0.05% trifluoroacetic acid); operating temperature: 64 ℃ C.; detector: 210nm)]。

Example 40

[8- (2-Acetylaminoethyl) -6, 7-dihydro-MeBmt]-1- [ (R) - (6-hydroxyhexyl) thio-Sar]-3-Cyclosporium sp Bacterins

6- (tosylthio) hexyl benzoate

P-tolylthiosulfonic acid potassium salt (18.00g, 79.52mmol) and 6-chloro-1-hexanol (10.86g, 79.52mmol) were added to ethanol (160 ml). The reaction mixture was stirred and heated to reflux for 5 hours. Most of the ethanol was evaporated under reduced pressure. The residue was mixed with ethyl acetate (200 ml). The ethyl acetate layer was washed with brine, dried over magnesium sulfate and evaporated to give 15.33g of crude S- (6-hydroxyhexyl) 4-methylthiobenzenesulfonate.

S- (6-hydroxyhexyl) 4-methylthiobenzenesulfonate (15.33g, 53.23mmol) was dissolved in acetone (60 ml). Benzoic anhydride (24.08, 106.46mmol), 4- (dimethylamino) pyridine (1.30g, 10.65mmol) and pyridine (30ml) were added. The mixture was stirred at room temperature overnight. Most of the acetone and pyridine were evaporated under reduced pressure. Ethyl acetate (100ml) was added. The ethyl acetate layer was washed with a hydrochloric acid solution (1.00N), brine, saturated sodium bicarbonate solution and brine, dried over magnesium sulfate and evaporated under reduced pressure. The residue was purified by chromatography (hexane/acetone) to give 5.41g of pure 6- (tosylthio) hexyl benzoate [ formula: c₂₀H₂₄O₄S₂(ii) a Accurate quality: 392.11, respectively; MS (M/z) 392.57(M +1)⁺(ii) a HPLC RT: 20.86 min. (C8 reverse phase column: 250 mm; acetonitrile/water (0.05% trifluoroacetic acid); operating temperature: 64 ℃ C.; detector: 210nm)]。

[8- (2- (tert-Butoxycarbonyl) aminoethyl) -6, 7-dihydro-MeBmt ] -1- [ (R) - (6- (benzoyloxy) hexyl) thio-Sar ] -3-cyclosporin

N-butyllithium (2.60M, 6.28ml, 16.33mmol) was added to a solution of diisopropylamine (2.32ml, 16.33mmol) in tetrahydrofuran (50ml) at-78 ℃ under nitrogen. After stirring the reaction mixture for 1 hour, [8- (2- (tert-butoxycarbonyl) aminoethyl) was added over 10 minutes-6, 7-dihydro-MeBmt]-1-Cyclosporin (2.20g, 1.63mmol) in tetrahydrofuran (15 ml). The mixture was stirred at-78 ℃ for 2 hours. A solution of 6- (tosylthio) hexyl benzoate (3.83g, 9.78mmol) in tetrahydrofuran (10ml) was added and the mixture was stirred at-78 ℃ for a further 2 h. The cooling bath was then removed and the reaction mixture was allowed to warm to room temperature and stirred for an additional 2 hours. Saturated ammonium chloride solution (20ml) was added to quench the reaction. Most of the tetrahydrofuran was removed under reduced pressure. Ethyl acetate (50ml) and brine (50ml) were added and the mixture was separated. The organic layer was dried over magnesium sulfate and evaporated under reduced pressure to give crude [8- (2- (tert-butoxycarbonyl) aminoethyl) -6, 7-dihydro-MeBmt]-1- [ (R) - (6- (benzoyloxy) hexyl) thio-Sar]-3-cyclosporin [ formula: c₈₂H₁₄₂N₁₂O₁₆S; accurate quality: 1583.04, respectively; MS (M/z) 1583.75(M +1)⁺(ii) a HPLC RT: 19.27 min. (C8 reverse phase column: 250 mm; acetonitrile/water (0.05% trifluoroacetic acid); operating temperature: 64 ℃ C.; detector: 210nm)]。

[8- (2- (tert-Butoxycarbonyl) aminoethyl) -6, 7-dihydro-MeBmt ] -1- [ (R) - (6-hydroxyhexyl) thio-Sar ] -3-cyclosporin

Reacting [8- (2- (tert-butoxycarbonyl) aminoethyl) -6, 7-dihydro-MeBmt]-1- [ (R) - (6- (benzoyloxy) hexyl) thio-Sar]-3-Cyclosporin was dissolved in methanol (10 ml). Lithium hydroxide (1.25g, 52.08mmol) and water (10ml) were added. The mixture was stirred at room temperature for 2 hours. Most of the methanol was then evaporated under reduced pressure. Ethyl acetate (50ml) and brine (50ml) were added and separated. The ethyl acetate layer was dried over magnesium sulfate and evaporated under reduced pressure. The residue was purified by chromatography (hexane/acetone) to give 0.34g of pure [8- (2- (tert-butoxycarbonyl) aminoethyl) -6, 7-dihydro-MeBmt]-1- [ (R) - (6-hydroxyhexyl) thio-Sar]-3-cyclosporin [ formula: c₇₅H₁₃₈N₁₂O₁₅S; accurate quality: 1479.01, respectively; MS (M/z) 1479.86(M +1)⁺,1501.80(M+Na)⁺；HPLC RT：17.86min。(C8And (3) reversed-phase column: 250 mm; acetonitrile/water (0.05% trifluoroacetic acid); operating temperature: 64 ℃; a detector: 210nm)]。

[8- (2-aminoethyl) -6, 7-dihydro-MeBmt ] -1- [ (R) - (6-hydroxyhexyl) thio-Sar ] -3-cyclosporin

Reacting [8- (2- (tert-butoxycarbonyl) aminoethyl) -6, 7-dihydro-MeBmt]-1- [ (R) - (6-hydroxyhexyl) thio-Sar]-3-Cyclosporin (0.34g, 0.15mmol) was dissolved in dichloromethane (15ml) and placed in an ice-water bath. Trifluoroacetic acid (5ml) was added. The mixture was stirred at 0 ℃ for three hours. Another portion of dichloromethane (50ml) was added. The dichloromethane layer was washed with brine (50ml), saturated sodium bicarbonate solution (50ml), dried over magnesium sulfate and evaporated under reduced pressure to give 0.14g of crude [8- (2-aminoethyl) -6, 7-dihydro-MeBmt]-1- [ (R) - (6-hydroxyhexyl) thio-Sar]-3-cyclosporin [ formula: c₇₀H₁₃₀N₁₂O₁₃S; accurate quality: 1378.96, respectively; MS (M/z) 1379.67(M +1)⁺(ii) a HPLC RT: 12.45 min. (C8 reverse phase column: 250 mm; acetonitrile/water (0.05% trifluoroacetic acid); operating temperature: 64 ℃ C.; detector: 210nm)]。

[8- (2-Acetylaminoethyl) -6, 7-dihydro-MeBmt ] -1- [ (R) - (6-hydroxyhexyl) thio-Sar ] -3-Cyclosporin

Crude [8- (2-aminoethyl) -6, 7-dihydro-MeBmt]-1- [ (R) - (6-hydroxyhexyl) thio-Sar]-3-Cyclosporin was dissolved in dichloromethane (15 ml). Acetic acid (0.14g, 2.43mmol), HBTU (0.55g, 1.46mmol), 1-hydroxybenzotriazole (0.20g, 1.46mmol) and pyridine (1.0ml) were added. The mixture was stirred at room temperature overnight. Most of the solvent was then evaporated under reduced pressure. Ethyl acetate (50ml) and brine (50ml) were added and separated. The ethyl acetate layer was dried over magnesium sulfate and evaporated under reduced pressure. By chromatographyThe residue was purified by method (hexane/acetone) to give 14mg of pure [8- (2-acetamidomethyl) -6, 7-dihydro-MeBmt]-1- [ (R) - (6-hydroxyhexyl) thio-Sar]-3-cyclosporin [ formula: c₇₂H₁₃₂N₁₂O₁₄S; accurate quality: 1420.97, respectively; MS (M/z) 1422.08(M +1)⁺,1443.93(M+Na)⁺(ii) a HPLC RT: 16.23 min. (C8 reverse phase column: 250 mm; acetonitrile/water (0.05% trifluoroacetic acid); operating temperature: 64 ℃ C.; detector: 210nm)]。

EXAMPLE 41

[8- (2-Acetylaminoethyl) -6, 7-dihydro-MeBmt]-1- [ (S) - ((4-hydroxy-2-methylbutan-2-yl) Thio) methyl-Sar]-3-Cyclosporin

3-methyl-3-mercaptobutanoic acid

3-hydroxy-3-methylbutyric acid (10.00g, 84.65mmol), hydrochloric acid (36%, 30ml) and thiourea (7.08g, 93.12mmol) were combined and heated at reflux for 24 h. The mixture was then evaporated under reduced pressure. Methanol (10ml) and a solution of lithium hydroxide (6.09g, 253.95mmol) in water (10ml) were added. The mixture was stirred and heated to reflux overnight. After cooling to room temperature, the mixture was filtered. The filtrate was evaporated under reduced pressure to give the crude product.

[8- (2- (tert-Butoxycarbonyl) aminoethyl) -6, 7-dihydro-MeBmt ] -1- [ (S) - ((1-carboxy-2-methylpropan-2-yl) thio) methyl-Sar ] -3-cyclosporin

To [8- (2- (tert-butoxycarbonyl) aminoethyl) -6, 7-dihydro-MeBmt]-1- [ α -methylene-Sar]3-methyl-3-mercaptobutanoic acid (0.30g, 2.20mmol) and hydrogen were added to a solution of (1.00g, 0.74mmol) of (E) -3-cyclosporin in methanol (10ml)Lithium oxide (0.18g, 7.36 mmol). The reaction mixture was stirred at room temperature overnight. Most of the methanol was evaporated under reduced pressure. Ethyl acetate (50ml) and brine (50ml) were added, and the pH of the aqueous layer was adjusted to 3 by the addition of hydrochloric acid solution (1.0N). The ethyl acetate layer was dried over magnesium sulfate and evaporated under reduced pressure to give crude [8- (2- (tert-butoxycarbonyl) aminoethyl) -6, 7-dihydro-MeBmt]-1- [ (S) - ((1-carboxy-2-methylpropan-2-yl) thio) methyl-Sar]-3-cyclosporin [ formula: c₇₅H₁₃₆N₁₂O₁₆S; accurate quality: 1492.99, respectively; MS (M/z) 1493.63(M +1)⁺,1515.88(M+Na)⁺]。

[8- (2- (tert-Butoxycarbonyl) aminoethyl) -6, 7-dihydro-MeBmt ] -1- [ (S) - ((4-methoxy-2-methyl-4-oxobutan-2-yl) thio) methyl-Sar ] -3-cyclosporin

The crude [8- (2- (tert-butoxycarbonyl) aminoethyl) -6, 7-dihydro-MeBmt]-1- [ (S) - ((1-carboxy-2-methylpropan-2-yl) thio) methyl-Sar]-3-Cyclosporin was dissolved in acetone (10 ml). Methyl iodide (0.14ml, 2.22mmol) and potassium carbonate (0.31g, 2.22mmol) were added. The mixture was stirred at room temperature for 2 hours. Most of the acetone was evaporated under reduced pressure. Ethyl acetate (50ml) and water (50ml) were then added and the mixture was separated. The ethyl acetate layer was dried over magnesium sulfate and evaporated under reduced pressure. The residue was purified by chromatography (hexane/acetone) to give 0.16g of pure [8- (2- (tert-butoxycarbonyl) aminoethyl) -6, 7-dihydro-MeBmt]-1- [ (S) - ((4-methoxy-2-methyl-4-oxobutan-2-yl) thio) methyl-Sar]-3-cyclosporin [ formula: c₇₆H₁₃₈N₁₂O₁₆S; accurate quality: 1507.01, respectively; MS (M/z) 1507.65(M +1)⁺,1529.95(M+Na)⁺(ii) a HPLC RT: 18.34 min. (C8 reverse phase column: 250 mm; acetonitrile/water (0.05% trifluoroacetic acid); operating temperature: 64 ℃ C.; detector: 210nm)]。

[8- (2-aminoethyl) -6, 7-dihydro-MeBmt ] -1- [ (S) - ((4-methoxy-2-methyl-4-oxobutan-2-yl) thio) methyl-Sar ] -3-cyclosporin

Reacting [8- (2- (tert-butoxycarbonyl) aminoethyl) -6, 7-dihydro-MeBmt]-1- [ (S) - (4-methoxy-2-methyl-4-oxobutan-2-ylthio) methyl-Sar]-3-Cyclosporin (0.16g, 0.11mmol) was dissolved in dichloromethane (6ml) and placed in an ice-water bath. Trifluoroacetic acid (2ml) was added. The mixture was stirred at 0 ℃ for three hours. Another portion of dichloromethane (20ml) was added. The mixture was washed with brine (30ml), saturated sodium bicarbonate solution (30ml), dried over magnesium sulfate and evaporated under reduced pressure to give crude [8- (2-aminoethyl) -6, 7-dihydro-MeBmt]-1- [ (S) - ((4-methoxy-2-methyl-4-oxobutan-2-yl) thio) methyl-Sar]-3-cyclosporin. The molecular formula is as follows: c₇₁H₁₃₀N₁₂O₁₄S; accurate quality: 1406.96, respectively; MS (M/z) 1407.65(M +1)⁺(ii) a HPLC RT: 13.66 min. (C8 reverse phase column: 250 mm; acetonitrile/water (0.05% trifluoroacetic acid); operating temperature: 64 ℃ C.; detector: 210nm)]。

[8- (2-Acetylaminoethyl) -6, 7-dihydro-MeBmt ] -1- [ (S) - ((4-methoxy-2-methyl-4-oxobutan-2-yl) thio) methyl-Sar ] -3-cyclosporin

Crude [8- (2-aminoethyl) -6, 7-dihydro-MeBmt]-1- [ (S) - ((4-methoxy-2-methyl-4-oxobutan-2-yl) thio) methyl-Sar]-3-Cyclosporin was dissolved in dichloromethane (10 ml). Acetic acid (0.03ml, 20.53mmol), HBTU (0.12g, 0.32mmol), 1-hydroxybenzotriazole (0.04g, 0.32mmol) and pyridine (0.50ml) were added. The mixture was stirred at room temperature for 2 hours. Dichloromethane (30ml) and brine (50ml) were then added and separated. The dichloromethane layer was dried over magnesium sulfate and evaporated under reduced pressure to give crude [8- (2-acetamidomethyl) -6, 7-dihydro-MeBmt]-1- [ (S) - ((4-methoxy-2-methyl-4-oxobutan-2-yl) thio) methyl-Sar]-3-cyclosporin [ formula: c₇₃H₁₃₂N₁₂O₁₅S; accurate quality: 1448.97, respectively; MS (M/z) 1449.72(M +1)⁺,1471.91(M+Na)⁺(ii) a HPLC RT: 17.19 min. (C8 reverse phase column: 250 mm; acetonitrile/water (0.05% trifluoroacetic acid); operating temperature: 64 ℃ C.; detector: 210nm)]。

[8- (2-Acetylaminoethyl) -6, 7-dihydro-MeBmt ] -1- [ (S) - ((4-hydroxy-2-methylbutan-2-yl) thio) methyl-Sar ] -3-Cyclosporin

Crude [8- (2-acetamidoethyl) -6, 7-dihydro-MeBmt]-1- [ (S) - ((4-methoxy-2-methyl-4-oxobutan-2-yl) thio) methyl-Sar]-3-Cyclosporin was dissolved in tetrahydrofuran (10 ml). Sodium borohydride (0.50g, 12.88mmol) and cesium chloride (0.10g, 0.59mmol) were added. The mixture was stirred at room temperature, and methanol (10ml) was added dropwise over 2 hours. The mixture was then stirred overnight. Most of the solvent was evaporated under reduced pressure. Ethyl acetate (50ml) and brine (50ml) were added and the mixture was separated. The ethyl acetate layer was dried over magnesium sulfate and evaporated under reduced pressure. The residue was purified by chromatography (dichloromethane/methanol) to give 34mg of pure [8- (2-acetamidomethyl) -6, 7-dihydro-MeBmt]-1- [ (S) - ((4-hydroxy-2-methylbutan-2-yl) thio) methyl-Sar]-3-cyclosporin [ formula: c₇₂H₁₃₂N₁₂O₁₄S; accurate quality: 1420.97, respectively; MS (M/z) 1421.87(M +1)⁺,1444.00(M+Na)⁺(ii) a HPLC RT: 15.67 min. (C8 reverse phase column: 250 mm; acetonitrile/water (0.05% trifluoroacetic acid); operating temperature: 64 ℃ C.; detector: 210nm)]。

Example 42

[8- (2-Acetylaminoethyl) -6, 7-dihydro-MeBmt]-1- [ (S) - ((5-hydroxy-2-methylpentane-2-yl) Thio) methyl-Sar]-3-Cyclosporin

4-mercapto-4-methylpentanoic acid

Methyl levulinate (10.00g, 76.84mmol) was dissolved in anhydrous tetrahydrofuran (50ml) under nitrogen. The mixture was placed in a dry ice-acetone bath and methyl magnesium chloride (3.00M, 25.6ml, 76.84mmol) was added slowly. The mixture was stirred at-78 ℃ for 2 hours and allowed to warm to room temperature overnight. Saturated ammonium chloride solution (20ml) was added to quench the reaction. Most of the tetrahydrofuran was removed under reduced pressure. Ethyl acetate (50ml) and brine (50ml) were added and the mixture was separated. The organic layer was dried over magnesium sulfate and evaporated under reduced pressure. The residue was purified by chromatography (hexane/acetone) to give 8.13g of pure methyl 4-hydroxy-4-methylpentanoate [ formula: c₇H₁₄O₃(ii) a Accurate quality: 146.09, respectively; MS (M/z) 146.64(M +1)⁺]。

Methyl 4-hydroxy-4-methylpentanoate (8.13g, 55.70mmol), hydrochloric acid (36%, 25ml) and thiourea (4.66g, 61.27mmol) were mixed and heated under reflux for 24 hours. The mixture was then evaporated under reduced pressure. Methanol (10ml) and a solution of lithium hydroxide (4.01g, 167.09mmol) in water (10ml) were added. The mixture was stirred and heated to reflux overnight. After cooling to room temperature, the mixture was filtered. The filtrate was evaporated under reduced pressure to give the crude product.

[8- (2- (tert-Butoxycarbonyl) aminoethyl) -6, 7-dihydro-MeBmt ] -1- [ (S) - ((4-carboxy-2-methylbutan-2-yl) thio) methyl-Sar ] -3-Cyclosporin

To [8- (2- (tert-butoxycarbonyl) aminoethyl) -6, 7-dihydro-MeBmt]-1- [ α -methylene-Sar]To a solution of (1.91g, 1.41mmol) of (E) -3-cyclosporin in methanol (20ml) were added (0.62g, 4.22mmol) of 4-mercapto-4-methylpentanoic acid and (0.20g, 8.43mmol) of lithium hydroxide. The reaction mixture was stirred at room temperatureStirring for two days. Most of the methanol was evaporated under reduced pressure. Ethyl acetate (100ml) and brine (100ml) were added and the pH of the aqueous layer was adjusted to 3 by the addition of hydrochloric acid solution (1.00N). After separation, the ethyl acetate layer was dried over magnesium sulfate and evaporated under reduced pressure to give crude [8- (2- (tert-butoxycarbonyl) aminoethyl) -6, 7-dihydro-MeBmt]-1- [ (S) - ((4-carboxy-2-methylbutan-2-yl) thio) methyl-Sar]-3-cyclosporin [ formula: c₇₆H₁₃₈N₁₂O₁₆S; accurate quality: 1507.01, respectively; MS (M/z) 1507.65(M +1)⁺,1529.95(M+Na)⁺]。

[8- (2- (tert-Butoxycarbonyl) aminoethyl) -6, 7-dihydro-MeBmt ] -1- [ (S) - ((5-methoxy-2-methyl-5-oxopentan-2-yl) thio) methyl-Sar ] -3-cyclosporin

The crude [8- (2- (tert-butoxycarbonyl) aminoethyl) -6, 7-dihydro-MeBmt]-1- [ (S) - ((4-carboxy-2-methylbutan-2-yl) thio) methyl-Sar]-3-Cyclosporin was dissolved in acetone (20 ml). Methyl iodide (0.56ml, 8.88mmol) and potassium carbonate (1.24g, 8.88mmol) were added. The mixture was stirred at room temperature over the weekend. Most of the acetone was evaporated under reduced pressure. Ethyl acetate (50ml) and water (50ml) were then added and the mixture was separated. The ethyl acetate layer was dried over magnesium sulfate and evaporated under reduced pressure. The residue was purified by chromatography (hexane/acetone) to give 0.45g of pure [8- (2- (tert-butoxycarbonyl) aminoethyl) -6, 7-dihydro-MeBmt]-1- [ (S) - ((5-methoxy-2-methyl-5-oxopentan-2-yl) thio) methyl-Sar]-3-cyclosporin [ formula: c₇₇H₁₄₀N₁₂O₁₆S; accurate quality: 1521.02, respectively; MS (M/z) 1521.61(M +1)⁺,1543.72(M+Na)⁺(ii) a HPLC RT: 18.31 min. (C8 reverse phase column: 250 mm; acetonitrile/water (0.05% trifluoroacetic acid); operating temperature: 64 ℃ C.; detector: 210nm)]。

[8- (2-aminoethyl) -6, 7-dihydro-MeBmt ] -1- [ (S) - ((5-methoxy-2-methyl-5-oxopentan-2-yl) thio) methyl-Sar ] -3-Cyclosporin

Reacting [8- (2- (tert-butoxycarbonyl) aminoethyl) -6, 7-dihydro-MeBmt]-1- [ (S) - ((5-methoxy-2-methyl-5-oxopentan-2-yl) thio) methyl-Sar]-3-Cyclosporin (0.45g, 0.30mmol) was dissolved in dichloromethane (15ml) and placed in an ice-water bath. Trifluoroacetic acid (5ml) was added. The mixture was stirred at 0 ℃ for 2 hours. Another portion of dichloromethane (50ml) was added. The mixture was washed with brine (50ml), saturated sodium bicarbonate solution (50ml), dried over magnesium sulfate and evaporated under reduced pressure to give crude [8- (2-aminoethyl) -6, 7-dihydro-MeBmt]-1- [ (S) - ((5-methoxy-2-methyl-5-oxopentan-2-yl) thio) methyl-Sar]-3-cyclosporin [ formula: c₇₂H₁₃₂N₁₂O₁₄S; accurate quality: 1420.97, respectively; MS (M/z) 1421.67(M +1)⁺(ii) a HPLC RT: 12.97 min. (C8 reverse phase column: 250 mm; acetonitrile/water (0.05% trifluoroacetic acid); operating temperature: 64 ℃ C.; detector: 210nm)]。

[8- (2-Acetylaminoethyl) -6, 7-dihydro-MeBmt ] -1- [ (S) - ((5-methoxy-2-methyl-5-oxopentan-2-yl) thio) methyl-Sar ] -3-Cyclosporin

Crude [8- (2-aminoethyl) -6, 7-dihydro-MeBmt]-1- [ (S) - ((5-methoxy-2-methyl-5-oxopentan-2-yl) thio) methyl-Sar]-3-Cyclosporin was dissolved in dichloromethane (15 ml). Acetic acid (0.09g, 1.48mmol), HBTU (0.34g, 0.89mmol), 1-hydroxybenzotriazole (0.12g, 0.89mmol) and triethylamine (0.50ml) were added. The mixture was stirred at room temperature for 2 hours. Dichloromethane (50ml) and brine (50ml) were then added and separated. The dichloromethane layer was dried over magnesium sulfate and evaporated under reduced pressure. The residue was purified by chromatography (hexane/acetone) to give 0.20mg of pure [8- (2-acetamidomethyl) -6, 7-dihydro-MeBmt]-1- [ (S) - ((5-methoxy-2-methyl-5-oxopentane-2)-radical) thio) methyl-Sar]-3-cyclosporin [ formula: c₇₄H₁₃₄N₁₂O₁₅S; accurate quality: 1462.98, respectively; MS (M/z) 1463.74(M +1)⁺,1485.81(M+Na)⁺(ii) a HPLC RT: 17.11 min. (C8 reverse phase column: 250 mm; acetonitrile/water (0.05% trifluoroacetic acid); operating temperature: 64 ℃ C.; detector: 210nm)]。

[8- (2-Acetylaminoethyl) -6, 7-dihydro-MeBmt ] -1- [ (S) - ((5-hydroxy-2-methylpentane-2-yl) thio) methyl-Sar ] -3-Cyclosporin

Crude [8- (2-acetamidoethyl) -6, 7-dihydro-MeBmt]-1- [ (S) - ((5-methoxy-2-methyl-5-oxopentan-2-yl) thio) methyl-Sar]-3-Cyclosporin (0.20g, 0.14mmol) was dissolved in tetrahydrofuran (10 ml). Sodium borohydride (1.00g, 64.40mmol) and cesium chloride (0.20g, 0.59mmol) were added. The mixture was stirred at room temperature, and methanol (10ml) was added dropwise over 2 hours. The mixture was then stirred overnight. Most of the solvent was evaporated under reduced pressure. Ethyl acetate (50ml) and brine (50ml) were added and the mixture was separated. The ethyl acetate layer was dried over magnesium sulfate and evaporated under reduced pressure. The residue was purified by chromatography (dichloromethane/methanol) to give 34mg of pure [8- (2-acetamidomethyl) -6, 7-dihydro-MeBmt]-1- [ (S) - ((5-hydroxy-2-methylpentane-2-yl) thio) methyl-Sar]-3-cyclosporin [ formula: c₇₃H₁₃₄N₁₂O₁₄S; accurate quality: 1434.99, respectively; MS (M/z) 1435.86(M +1)⁺,1457.87(M+Na)⁺(ii) a HPLC RT: 17.03 min. (C8 reverse phase column: 250 mm; acetonitrile/water (0.05% trifluoroacetic acid); operating temperature: 64 ℃ C.; detector: 210nm)]。

Reference example 1

[8- (2-Acetylaminoethyl) -6, 7-dihydro-MeBmt]-1- [ (R) -2-methyl-Sar]-3-Cyclosporin (CPI-431-32(CRV431))

[ (R) -2-methyl-Sar ] -3-Cyclosporin

N-butyllithium (2.20M, 75.60ml, 166.39mmol) was added to a solution of diisopropylamine (23.60ml, 166.39mmol) in tetrahydrofuran (150ml) at-78 ℃ under nitrogen. After stirring the reaction mixture for 1 hour, a solution of cyclosporin (20.00g, 16.64mmol) in tetrahydrofuran (50ml) was added over 10 minutes. The mixture was stirred at-78 ℃ for 2 hours. After addition of iodomethane (10.36ml, 166.39mmol), the mixture was stirred at-78 ℃ for a further 2 hours and allowed to warm to room temperature overnight. Brine (20ml) was added to quench the reaction. Most of the tetrahydrofuran was removed under reduced pressure. Ethyl acetate (100ml) and brine (100ml) were added and the mixture was separated. The organic layer was dried over magnesium sulfate and evaporated under reduced pressure. The residue was purified by chromatography (hexane/acetone) to give 5.49g of [ (R) -2-methyl-Sar)]-3-cyclosporin [ formula: c₆₃H₁₁₃N₁₁O₁₂(ii) a Accurate quality: 1215.86, respectively; MS (M/z) 1216.63(M +1)⁺,1238.79(M+Na)⁺(ii) a HPLC RT: 17.53 min. (C8 reverse phase column: 250 mm; acetonitrile/water (0.05% trifluoroacetic acid); operating temperature: 64 ℃ C.; detector: 210nm)]。

[ 3-acetyl-MeBmt ] -1- [ (R) -2-methyl-Sar ] -3-Cyclosporin

To a dry flask was added [ (R) -2-methyl-Sar ] under nitrogen]-3-Cyclosporine (5.49g, 4.52mmol), N-dimethylaminopyridine (0.06g, 0.45mmol), anhydrous pyridine (60ml) and acetic anhydride (28.21ml, 299.00 mol). The reaction mixture was stirred at room temperature overnight. The mixture was poured into ice water (300ml) and stirred until the ice melted. Ethyl acetate (100ml) was added and the mixture was separated. The ethyl acetate layer was washed with 1N hydrochloric acid solution (50 ml. times.2), saturated sodium bicarbonate solution (50ml), brine (50ml) and dried over magnesium sulfateDried and evaporated under reduced pressure. The residue was purified by chromatography (hexane/acetone/methanol) to give 4.67g of [ 3-acetyl-MeBmt]-1- [ (R) -2-methyl-Sar]-3-cyclosporin [ formula: c₆₅H₁₁₅N₁₁O₁₃(ii) a Accurate quality: 1257.87, respectively; MS (M/z) 1258.54(M +1)⁺,1280.71(M+Na)⁺(ii) a HPLC RT: 19.31 min. C8 reverse phase column: 250 mm; acetonitrile/water (0.05% trifluoroacetic acid); operating temperature: 64 ℃; a detector: 210nm)]。

[ (3R,4R) -3-acetoxy-4-methyl-6-oxo-N-Menle ] -1- [ (R) -2-methyl-Sar ] -3-Cyclosporin

To [ 3-acetyl-MeBmt]-1- [ (R) -2-methyl-Sar]To a solution of-3-cyclosporin (4.67g, 3.71mmol) in dioxane (100ml) was added water (20ml), osmium (VIII) oxide solution (15.74mM, 23.50ml, 0.37mmol) and sodium metaperiodate (3.18g, 14.85 mmol). The reaction mixture was stirred at room temperature for 5 hours. Ethyl acetate (100ml) and brine (100ml) were added and the mixture was separated. The organic layer was washed with saturated sodium bicarbonate solution, brine, dried over magnesium sulfate and evaporated under reduced pressure. The residue was purified by chromatography (hexane/acetone) to give 4.10g of [ (3R,4R) -3-acetoxy-4-methyl-6-oxo-N-Menle]-1- [ (R) -2-methyl-Sar]-3-cyclosporin [ formula: c₆₃H₁₁₁N₁₁O₁₄(ii) a Accurate quality: 1245.83, respectively; MS (M/z) 1246.54(M +1)⁺,1268.71(M+Na)⁺(ii) a HPLC RT: 17.27 min. 8, reversed phase column: 250 mm; acetonitrile/water (0.05% trifluoroacetic acid); operating temperature: 64 ℃; a detector: 210nm)]。

[ 8-cyanomethyl-3-acetyl-MeBmt ] -1- [ (R) -2-methyl-Sar ] -3-cyclosporin

Under nitrogen toA dry flask was charged with (3-cyanopropyl) triphenyl phosphonium bromide (7.98g, 19.50mmol) and anhydrous tetrahydrofuran (60 ml). The reaction mixture was placed in an ice-water bath and sodium tert-butoxide (2.19g, 22.75mmol) was added. After stirring the mixture for 2 hours [ (3R,4R) -3-acetoxy-4-methyl-6-oxo-N-Menle was added]-1- [ (R) -2-methyl-Sar]-3-Cyclosporin (4.10g, 3.29mmol) in anhydrous tetrahydrofuran (20 ml). The mixture was stirred at 0 ℃ for a further 5 hours. Saturated ammonium chloride solution (20ml) was then added to quench the reaction. Most of the tetrahydrofuran was evaporated under reduced pressure. Ethyl acetate (100ml) and brine (100ml) were added and the mixture was separated. The organic layer was dried over magnesium sulfate and evaporated under reduced pressure. The residue was purified by chromatography (dichloromethane/methanol) to give 2.00g of pure [ 8-cyanomethyl-3-acetyl-MeBmt]-1- [ (R) -2-methyl-Sar]-3-cyclosporin [ formula: c₆₇H₁₁₆N₁₂O₁₃(ii) a Accurate quality: 1296.88, respectively; MS (M/z) 1297.55(M +1)⁺,1319.787(M+Na)⁺(ii) a HPLC RT: 17.56 min. C8 reverse phase column: 250 mm; acetonitrile/water (0.05% trifluoroacetic acid); operating temperature: 64 ℃; a detector: 210nm)]。

[8- (2-aminoethyl) -3-acetyl-6, 7-dihydro-MeBmt ] -1- [ (R) -2-methyl-Sar ] -3-Cyclosporin

To [ 8-cyanomethyl-3-acetyl-MeBmt under nitrogen]-1- [ (R) -2-methyl-Sar]To a solution of (2.00g, 1.54mmol) of (E) -3-cyclosporin in methanol (50ml) was added nickel (II) chloride hexahydrate (0.04g, 0.15 mmol). The reaction mixture was placed in an ice-water bath. Sodium borohydride (3.05g, 80.50mmol) was added in four portions over a two hour period. After the mixture was stirred at 0 ℃ for another 2 hours, water (10ml) was added. Most of the methanol was evaporated under reduced pressure. Ethyl acetate (50ml) and saturated sodium bicarbonate solution (50ml) were added to the solution, and the mixture was separated. The organic layer was washed with brine, dried over magnesium sulfate and evaporated under reduced pressure. The residue was dissolved in methanol (30 ml). Palladium (10 wt% on carbon, 20mg) and acetic acid (5 drops) were added. Mixing the mixture inStir at room temperature under hydrogen for 2 hours. The mixture was then filtered and the filtrate evaporated under reduced pressure to give 2.20g of crude [8- (2-aminoethyl) -3-acetyl-6, 7-dihydro-MeBmt]-1- [ (R) -2-methyl-Sar]-3-cyclosporin [ formula: c₆₇H₁₂₂N₁₂O₁₃(ii) a Accurate quality: 1302.93, respectively; MS (M/z) 1303.75(M +1)⁺(ii) a HPLC RT: 14.72 min. (C8 reverse phase column: 250 mm; acetonitrile/water (0.05% trifluoroacetic acid); operating temperature: 64 ℃ C.; detector: 210nm)]。

[8- (2-aminoethyl) -6, 7-dihydro-MeBmt ] -1- [ (R) -2-methyl-Sar ] -3-Cyclosporin

Reacting [8- (2-aminoethyl) -3-acetyl-6, 7-dihydro-MeBmt]-1- [ (R) -2-methyl-Sar]-3-Cyclosporin (1.60g,1.24mmol) was dissolved in methanol (20 ml). Water (10ml) and tetramethylammonium hydroxide pentahydrate (0.67g, 3.72mmol) were added. The mixture was stirred at room temperature for 2 hours. Most of the methanol was evaporated. Ethyl acetate (100ml) and brine (100ml) were added and the mixture was separated. The organic layer was dried over magnesium sulfate and evaporated under reduced pressure. The residue was purified by chromatography (dichloromethane/methanol) to give 1.20g of pure [8- (2-aminoethyl) -6, 7-dihydro-MeBmt]-1- [ (R) -2-methyl-Sar]-3-cyclosporin [ formula: c₆₅H₁₂₀N₁₂O₁₂(ii) a Accurate quality: 1260.91, respectively; MS (M/z) 1261.72(M +1)⁺(ii) a HPLCRT: 12.11 min. (C8 reverse phase column: 250 mm; acetonitrile/water (0.05% trifluoroacetic acid); operating temperature: 64 ℃ C.; detector: 210nm)]。

[8- (2-Acetylaminoethyl) -6, 7-dihydro-MeBmt ] -1- [ (R) -2-methyl-Sar ] -3-Cyclosporin

Reacting [8- (2-aminoethyl) -6, 7-dihydro-MeBmt]-1- [ (R) -2-methyl-Sar]-3-Cyclosporin(0.60g,0.48mmol) was dissolved in dichloromethane (25 ml). Acetic acid (0.14ml, 2.38mmol), HBTU (0.54g, 1.43mmol), 1-hydroxybenzotriazole (0.19g, 1.43mmol) and pyridine (1.00ml) were added. The mixture was stirred at room temperature for three hours. Dichloromethane (30ml) and brine (50ml) were then added and separated. The dichloromethane layer was dried over magnesium sulfate and evaporated under reduced pressure. The residue was purified by chromatography (dichloromethane/methanol) to give 0.11g of pure [8- (2-acetamidomethyl) -6, 7-dihydro-MeBmt]-1- [ (R) -2-methyl-Sar]-3-cyclosporin [ formula: c₆₇H₁₂₂N₁₂O₁₃(ii) a Accurate quality: 1302.93, respectively; MS (M/z) 1303.66(M +1)⁺,1325.86(M+Na)⁺(ii) a HPLC RT: 16.56 min. (C8 reverse phase column: 250 mm; acetonitrile/water (0.05% trifluoroacetic acid); operating temperature: 64 ℃ C.; detector: 210nm)]. The product was prepared using a method similar to that described in U.S. patent nos. 9,200,038B2 and US 2013/0190223a1, which are incorporated herein by reference.

Reference example 2

[8- (2-Acetylaminoethyl) -6, 7-dihydro-MeBmt]-1-Cyclosporin

Reacting [8- (2-aminoethyl) -6, 7-dihydro-MeBmt]-1-Cyclosporin (0.50g,0.40mmol) was dissolved in dichloromethane (20 ml). Acetic acid (0.11ml, 2.00mmol), HBTU (0.46g, 1.20mmol), 1-hydroxybenzotriazole (0.16g, 1.20mmol) and pyridine (0.50ml) were added. The mixture was stirred at room temperature for one hour. Dichloromethane (30ml) and brine (50ml) were then added and separated. The dichloromethane layer was dried over magnesium sulfate and evaporated under reduced pressure. The residue was purified by chromatography (dichloromethane/methanol) to give 0.20g of pure [8- (2-acetamidomethyl) -6, 7-dihydro-MeBmt]-1-cyclosporin [ formula: c₆₆H₁₂₀N₁₂O₁₃(ii) a Accurate quality: 1288.91, respectively; MS (M/z) 1289.96(M +1)⁺,1311.82(M+Na)⁺(ii) a HPLC RT: 14.92 min. C8 reverse phase column: 250 mm; acetonitrile/water (0.05% trisFluoroacetic acid); operating temperature: 64 ℃; a detector: 210nm)]。

Reference example 3

[ α -methylene-Sar]-3-Cyclosporin

[ α -Methoxycarbonyl-Sar ] -3-Cyclosporin

[ α -carboxy-sar]-3-Cyclosporin (5.00g, 4.01mmol) was dissolved in N, N-dimethylformamide (30 ml). Methyl iodide (2.85g, 20.10mmol) and potassium carbonate (1.38g, 10.00mmol) were added. The mixture was stirred at room temperature for 2 hours. Ethyl acetate (60ml) and water (60ml) were then added and the mixture was separated. The ethyl acetate layer was washed with brine, dried over magnesium sulfate and evaporated under reduced pressure to give 5.32g of crude product which was used directly in the next step without purification (yield: -100%) [ formula: c₆₄H₁₁₃N₁₁O₁₄(ii) a Accurate quality: 1259.85, respectively; MS (M/z) 1260.7(M +1)⁺,1282.7(M+Na)⁺]。

[ (R) - α -hydroxymethyl-Sar ] -3-cyclosporin

Reacting [ α -methoxycarbonyl-Sar]-3-Cyclosporin (2.00g,1.59mmol) was dissolved in tetrahydrofuran (30 ml). Cesium chloride (1.33g, 7.90mmol) and sodium borohydride (0.60g, 15.89mmol) were added. Methanol (30ml) was then added dropwise to the mixture over a period of 2 hours. After the addition, the mixture was stirred at room temperature overnight. Most of the solvent was then evaporated under reduced pressure. Ethyl acetate (50ml) and water (50ml) were added. The ethyl acetate layer was separated and washed with brine, dried over magnesium sulfate and evaporated under reduced pressure to give 1.99g of a crude product, which was purified through a silica gel column using methylene chloride/methanol (100:0 to 95:5) to give 1.50g of a pure product (yield: 76%) [ min ]The subformula: c₆₃H₁₁₃N₁₁O₁₃(ii) a Accurate quality: 1231.85, respectively; MS (m/z): 1232.7(M +1)⁺,1254.7(M+Na)⁺]。

[ α -Methylmethanesulfonate-Sar ] -3-Cyclosporin

To 0 ℃ of [ α -hydroxymethyl-Sar]To a solution of-3-cyclosporin (30mg, 0.024mmol) in dichloromethane (2ml) were added triethylamine (52.8. mu.l, 0.38mmol) and methanesulfonyl chloride (23mg, 0.20 mmol). After stirring at room temperature for two hours, the mixture was washed with brine, dried over magnesium sulfate and evaporated under reduced pressure to give 33mg of crude product, which was used directly in the next step reaction without further purification [ formula: c₆₄H₁₁₅N₁₁O₁₅S; accurate quality: 1309.83, respectively; MS (m/z): 1310.7(M +1)⁺]。

[ α -chloromethyl-Sar ] -3-cyclosporin

To 0 ℃ of [ α -hydroxymethyl-Sar]To a solution of-3-cyclosporin (30mg, 0.024mmol) in dichloromethane (2ml) were added triethylamine (52.8. mu.l, 0.384mmol, 16 equivalents) and methanesulfonyl chloride (23mg, 0.20 mmol). After stirring at room temperature overnight, the mixture was washed with brine, dried over magnesium sulfate and evaporated under reduced pressure to give 30mg of crude product, which was used directly in the next step reaction without further purification [ formula: c₆₃H₁₁₂ClN₁₁O₁₂(ii) a Accurate quality: 1249.82, respectively; MS (m/z): 1250.7(M +1)⁺,1272.9(M+Na)⁺]。

[ α -methylene-Sar ] -3-Cyclosporin

Method 1

To [ α -methanesulfonic acid methyl ester-Sar at 0 DEG C]-3-cyclosporin (33mg, 0.025mmol) or [ α -chloromethyl-Sar]To a solution of-3-cyclosporin (30mg, 0.025mmol) in tetrahydrofuran (3ml) was added sodium hydride (15.3mg, 60% in oil, 0.38mmol, 10 equivalents). The mixture was stirred at 0 ℃ for 1 hour, then warmed to room temperature for 30 minutes. After removal of the solvent, the residue was dissolved in dichloromethane (20 ml). The dichloromethane layer was washed with 1N hydrochloric acid, saturated sodium bicarbonate solution and brine, dried over magnesium sulfate and evaporated under reduced pressure. The residue was purified by chromatography on silica gel with dichloromethane/methanol (20/1) to give 16mg of product (yield: 54%) [ formula: c₆₃H₁₁₁N₁₁O₁₂(ii) a Accurate quality: 1213.84, respectively; MS (m/z): 1214.7(M +1)⁺,1236.7(M+Na)⁺；TLC R_f: 0.55 (ethyl acetate/methanol-20/1); HPLC RT: 7.0min (C8 reverse phase column: 150 mm; acetonitrile/water (0.05% trifluoroacetic acid); operating temperature: 64 ℃ C.; detector: 210nm)]。

[ α -methylene-Sar ] -3-Cyclosporin

Method 2

[ α -methylene-Sar ] -3-cyclosporine may also be prepared using a method similar to that described in WO2012/051194A1 (which is incorporated herein by reference).

Reference example 4

[ α -methylene-Sar]-3- [ (γ -hydroxy) -NMeLeu]-4-Cyclosporin

[ α -Methoxycarbonylsar-Sar ] -3- [ (γ -hydroxy) -NMeLeu ] -4-Cyclosporin

To a solution of LDA (2.0M in tetrahydrofuran, 23ml, 46mmol) in tetrahydrofuran (80ml) was added [ (gamma-hydroxy) -N-MeLeu ] under nitrogen over 3 minutes at-78 deg.C]-4-Cyclosporin (4.40g, 3.61mmol) in tetrahydrofuran (15 ml). After the mixture was stirred at-78 ℃ for 3 hours, carbon dioxide gas was bubbled into the reaction mixture for 1 hour. The mixture was then slowly warmed to room temperature and kept stirring for 3 hours. Most of the tetrahydrofuran was evaporated. Dichloromethane (100ml) and water (50ml) were added. The pH of the mixture was adjusted to about 5 by adding aqueous citric acid. The mixture was separated and the organic layer was washed with brine, dried over magnesium sulfate and evaporated under reduced pressure to give 3.20g of crude product as acid which was used in the next step without purification [ formula: c₆₃H₁₁₁N₁₁O₁₅(ii) a Accurate quality: 1261.83, respectively; MS (m/z): 1262.49(M +1)⁺]To [ α -carboxy-Sar ]]-3- [ (γ -hydroxy) -N-MeLeu]To a mixture of (3.20g, 2.53mmol) of (E) -4-cyclosporin and (1.30g, 9.40mmol) of potassium carbonate in N, N-dimethylformamide (20ml) was added methyl iodide (1.80g, 12.70 mmol). The mixture was stirred at room temperature overnight. Dichloromethane (80ml) and water (50ml) were added and the mixture was separated. The dichloromethane layer was washed with water (25ml) and brine (25ml), dried over magnesium sulphate and evaporated under reduced pressure to give 3.00g of product [ formula: c₆₄H₁₁₃N₁₁O₁₅(ii) a Accurate quality: 1275.84, respectively; MS (m/z): 1276.75(M +1)⁺]。

[ (gamma-hydroxy) -N-MeLeu ] -4-cyclosporin is prepared by Sebekia benihana Biotransformation according to the method described by Kuhnt M. et al, 1996, Microbial Biotransformation Products of Cyclosporin A, J.antibiotics,49(8), 781.

[ (R) - α -hydroxymethyl-Sar ] -3- [ (γ -hydroxy) -NMeLeu ] -4-cyclosporin

To [ α -methoxycarbonyl-Sar ]]-3- [ (γ -hydroxy) -N-MeLeu]To a suspension of-4-cyclosporin (3.00g, 2.35mmol) and lithium chloride (1.50g, 35.30mmol) in methanol (100ml) was added sodium borohydride (2.50g, 66.10mmol) portionwise. The mixture was stirred at room temperature overnight. Most of the solvent was evaporated under reduced pressure. Dichloromethane (80ml) and water (50ml) were added and the mixture was separated. The dichloromethane layer was washed with brine, dried over magnesium sulfate and evaporated under reduced pressure. The residue was purified by chromatography (dichloromethane/methanol ═ 96/4) to give 1.30g of product [ formula: c₆₃H₁₁₃N₁₁O₁₄(ii) a Accurate quality: 1247.85, respectively; MS (m/z): 1248.48(M +1)⁺；¹H NMR Spectrum (600MHz, CDCl)₃δ in ppm): 0.68(d, J ═ 5.4Hz,3H),0.80-1.00(m,30H),1.07(d, J ═ 6.0Hz,3H), 1.16-1.29 (m,10H),1.32(d, J ═ 7.2Hz,3H),1.39-1.46(m,2H),1.59-1.63(m,6H),1.68-1.83(m,7H),2.02-2.11(m,4H),2.31-2.33(m,1H),2.37-2.42(m,2H),2.67(s,6H),3.09(s,3H),3.19(s,3H),3.20(s,3H),3.22(s,3H),3.47(s,3H), 3.72-3.72 (s,3H), 3.81 (m,3H), 3.65 (m,4H), 3.7-4H), 3.79 (m,4H), 3.7H, 3.7 (m, 3.7H), 1H) 4.90-4.95(m,2H), 5.00-5.05 (m,2H),5.09(d, J ═ 10.8Hz,1H),5.30-5.35(m,2H),5.46(d, J ═ 6.0Hz,1H),5.52-5.53(m,1H),5.66-5.68(m,1H),7.12(d, J ═ 7.8Hz,1H),7.47(d, J ═ 8.4Hz,1H),7.60(d, J ═ 7.2Hz,1H),7.87-7.89(d, J ═ 9.6Hz,1H)]。

[ α -methylene-Sar ] -3- [ (γ -hydroxy) -NMeLeu ] -4-Cyclosporin

Method 1

At room temperature to [ α -hydroxymethyl-Sar]-3- [ (γ -hydroxy) -NMeLeu]To a solution of (0.25g, 0.20mmol) of (4-cyclosporin in) dichloromethane (10ml) were added triethylamine (0.33ml, d 0.726, 2.40mmol) and triethylamine hydrochloride (95.6mg, 1.00mmol), followed by addition of p-toluenesulfonyl chloride (0.23g, 1.20mmol) with stirring. The mixture was stirred at room temperatureThe reaction mixture was then washed with brine, dried over magnesium sulfate and the solvent was evaporated under reduced pressure [ α -chloromethyl-Sar ]]-3- [ (γ -hydroxy) -NMeLeu]-4-cyclosporin [ formula: c₆₃H₁₁₂ClN₁₁O₁₃(ii) a Accurate quality: 1265.81, respectively; MS (m/z): 1266.32(M +1)⁺,1288.43(M+Na)⁺]And [ α -p-toluenesulfonylmethyl-Sar]-3- [ (γ -hydroxy) -NMeLeu]-4-cyclosporin [ formula: c₇₀H₁₁₉N₁₁O₁₆S; accurate quality: 1401.856, respectively; MS (m/z): 1402.34(M +1)⁺,1424.62(M+Na)⁺]The reaction mixture of (a) was used directly in the next step without further purification. To a solution of the above mixture in tetrahydrofuran (20ml) was added sodium hydride (320mg, 60% in oil, 8mmol) at 0 ℃. The mixture was stirred at 0 ℃ for one hour and then warmed to room temperature for 30 minutes. The reaction was quenched with saturated ammonium chloride solution. After removal of the tetrahydrofuran, the crude product was extracted with ethyl acetate. The ethyl acetate layer was washed with brine, dried over magnesium sulfate and evaporated under reduced pressure. The residue was purified by chromatography on silica gel using ethyl acetate/methanol (20/1) to give 45mg of the product (yield: 18%) [ formula: c₆₃H₁₁₁N₁₁O₁₃(ii) a Accurate quality: 1229.84, respectively; MS (m/z): 1230.6(M +1)⁺,1252.82(M+Na)⁺；TLC R_f: 0.50 (ethyl acetate/methanol-10/1); HPLC RT:15.38min (C8 reverse phase column: 250 mm; acetonitrile/water (0.05% trifluoroacetic acid); operating temperature: 64 ℃ C.; detector: 210 nm);¹h NMR Spectrum (600MHz, CDCl)₃δ in ppm): 0.72(d, J ═ 5.4Hz,3H),0.84-1.00(m,30H),1.17-1.26(m,15H),1.34(d, J ═ 6.0Hz,3H), 1.44-1.47 (m,2H),1.59-1.62(m,6H),1.69-1.76(m,4H),1.94-1.99(m,1H),2.09-2.13(m,3H),2.34-2.37(m,3H),2.65(s,3H),2.67(s,3H),3.09(s,3H)),3.10(s,3H),3.19(s,3H),3.44(s,3H),3.46(s,3H),3.80(m,1H),3.91(m, 4H), 4.47 (m, 4.81H), 4.47 (m,4H),1.9, 4H), 4.9-1.4H), 1.81 (m,4H), 1.9-1.4H), 1.9, 4H), 5.24(s,1H),5.32(m,2H),5.41-5.43(m,2H),5.64-5.66(m,1H),7.11(d, J ═ 7.2Hz,1H),7.49(d, J ═ 7.2Hz,1H),7.74(d, J ═ 8.4Hz,1H),7.84(d, J ═ 9.6Hz,1H)]。

[ α -methylene-Sar ] -3- [ (γ -hydroxy) -NMeLeu ] -4-Cyclosporin

Method 2

Mixing [ (R) - α -hydroxymethyl-Sar]-3- [ (γ -hydroxy) -NMeLeu]-4-Cyclosporin (crude, 2.00g), carbon tetrabromide (2.66g, 8.02mmol) and triphenylphosphine (2.11g, 8.02mmol) are dissolved in dichloromethane (30 ml.) the mixture is stirred at room temperature under nitrogen for two hours then the mixture is added under nitrogen at 0 ℃ to a suspension of sodium hydride (60% dispersed in mineral oil) (0.77g, 19.25mmol) in tetrahydrofuran (30 ml.) the mixture is stirred at 0 ℃ for one hour then most of the solvent is evaporated under reduced pressure the residue is slowly treated with water (10ml) at 0 ℃, ethyl acetate (30ml) and water (30ml) are added and the mixture is separated the ethyl acetate layer is washed with brine, dried over magnesium sulfate and evaporated under reduced pressure the residue is purified by chromatography (hexane/acetone from 90/10 to 70/30) to yield 0.68g of the product [ α -methylene-r]-3- [ (γ -hydroxy) -NMeLeu]-4-cyclosporin [ formula: c₆₃H₁₁₁N₁₁O₁₃(ii) a Accurate quality: 1229.84, respectively; MS (m/z): 1230.50(M +1)⁺,1252.68(M+Na)⁺；TLC R_f: 0.50 (ethyl acetate/methanol-10/1); HPLC RT: 15.36min (C8 reverse phase column: 250 mm; acetonitrile/water (0.05% trifluoroacetic acid); operating temperature: 64 ℃ C.; detector: 210nm)]。

[ α -methylene-Sar ] -3- [ (γ -hydroxy) -NMeLeu ] -4-Cyclosporin

Method 3

To [ (R) - α -hydroxymethyl-Sar under nitrogen at 0 deg.C]-3- [ (γ -hydroxy) -NMeLeu]-4-Cyclosporin (0.25g, 0.20mmol) in dichloromethane (10)ml) 1-chloro-N, N, 2-trimethyl-1-propenamine (131. mu.l, d1.01, 1.0mmol) was added dropwise to the solution, after stirring at 0 ℃ for 30 minutes, the mixture was warmed to room temperature and stirred for an additional hour, the reaction mixture was washed with sodium bicarbonate solution, brine, dried over magnesium sulphate and evaporated under reduced pressure, containing [ α -chloromethyl-Sar]-3- [ (γ -hydroxy) -NMeLeu]-4-cyclosporin [ formula: c₆₃H₁₁₂ClN₁₁O₁₃(ii) a Accurate quality: 1265.81, respectively; MS (m/z): 1266.32(M +1)⁺,1288.43(M+Na)⁺]The crude product of (a) was used in the next step without further purification. To a solution of the above crude product in tetrahydrofuran (20ml) was added sodium hydride (320mg, 60% in oil, 8mmol) with stirring at 0 ℃. The mixture was stirred at 0 ℃ for one hour and then warmed to room temperature for an additional 30 minutes. The reaction was then quenched with saturated ammonium chloride solution. After removal of the tetrahydrofuran, the residue was extracted with ethyl acetate. The ethyl acetate layer was washed with brine, dried over magnesium sulfate and evaporated under reduced pressure. The residue was purified by chromatography on silica gel with ethyl acetate/methanol (20/1) to give 33mg of product (yield: 13%) [ formula: c₆₃H₁₁₁N₁₁O₁₃(ii) a Accurate quality: 1229.84, respectively; MS (m/z): 1230.45(M +1)⁺,1252.65(M+Na)⁺；TLC R_f: 0.50 (ethyl acetate/methanol-10/1); HPLC RT: 15.36min (C8 reverse phase column: 250 mm; acetonitrile/water (0.05% trifluoroacetic acid); operating temperature: 64 ℃ C.; detector: 210nm)]。

[ α -methylene-Sar ] -3- [ (γ -hydroxy) -NMeLeu ] -4-Cyclosporin

Method 4

N-butyllithium (2.2M, 49.30ml, 108.46mmol) was added to a solution of diisopropylamine (15.39ml, 108.46mmol) in tetrahydrofuran (150ml) at-78 ℃ under nitrogen. After stirring the reaction mixture for 1 hour, [ (gamma-hydroxy) -NMeLeu was added over 10 minutes]-4-Cyclosporin(12.00g, 9.86mmol) of tetrahydrofuran (30ml) solution stirring was continued for two hours at-78 deg.C carbon dioxide gas was bubbled through the reaction mixture for two hours and the mixture was stirred for an additional hour at-78 deg.C then the cooling bath was removed and the reaction mixture was slowly warmed to room temperature with bubbling out unreacted carbon dioxide, the mixture was cooled to about 0-5 deg.C by an ice bath and chloromethyl chloroformate (13.98g, 108.46mmol) was added, the mixture was warmed to room temperature and stirred overnight, water (30ml) was added to quench the reaction then most of the solvent was evaporated under reduced pressure ethyl acetate (100ml) and water (80ml) were added, the ethyl acetate layer was separated and washed with brine, dried over magnesium sulfate and evaporated under reduced pressure hexane/acetone (from 90:10 to 70:30) was used as eluent to purify the residue by chromatography to give 4.74g of pure product [ α -methylene-r 30g SaSaSaSaSa]-3- [ (γ -hydroxy) -NMeLeu]-4-cyclosporin [ formula: c₆₃H₁₁₁N₁₁O₁₃(ii) a Accurate quality: 1229.84, respectively; MS (m/z): 1230.39(M +1)⁺,152.59(M+Na)⁺；TLC R_f: 0.50 (ethyl acetate/methanol-10/1); HPLC RT:15.38min (C8 reverse phase column: 250 mm; acetonitrile/water (0.05% trifluoroacetic acid); operating temperature: 64 ℃ C.; detector: 210nm)]The [ α -methylene-Sar was prepared using a procedure analogous to that described in WO2012/051194a1]-3- [ (γ -hydroxy) -NMeLeu]-4-cyclosporin.

Example 43

anti-HCV activity of cyclosporin derivatives

Cyclosporine derivatives were evaluated for anti-HCV activity in an HCV subgenomic replicon assay. This assay used the cell line ET (Luc-ubi-neo/ET), a Huh7 human hepatoma cell line carrying HCV replicons with a stable luciferase (Luc) reporter. HCV RNA replication was assessed by quantifying HCV replicon-derived luciferase activity. Evaluation of antiviral Activity of Cyclosporin analogs following drug treatment to deduce EC by use of luciferase endpoint₅₀And EC₉₀Values (Krieger, N., et al, 2001, J.Virol.7)5,4614-4624; pietschmann, T., et al, 2002, J.Virol.76, 4008-4021; each of which is incorporated herein by reference). Cytotoxicity was evaluated in parallel.

TABLE 2 test results for certain representative compounds

Antiviral activity: IC₅₀<35nM；***35nM<IC₅₀<90nM；**90nM<IC₅₀<250nM；*250nM<IC₅₀<450 nM; is inactive>1500nM。

Example 44

Cyclophilin a and D (CyPA and CyPD) PPI enzyme inhibition assay

Human recombinant CyPA (atgen) was dissolved in isomerase buffer (50mM Hepes, 100mM NaCl, 1mg/ml bovine serum albumin, 1mg/ml α -chymotrypsin; pH 8) to 10 nM. succinyl-AAPF-pNA peptide substrate (Sigma) was dissolved in dry LiCl/trifluoroethanol to 3.2 mM. each test compound prepared at 10 concentrations in DMSO, then diluted into CyPA-isomerase buffer to 0.05-1000nM (reaction mixture). equilibrating all solutions and performing the reaction at 5 ℃. the reaction was initiated by mixing 95 μ L of reaction mixture with 5 μ L of peptide pre-loaded in a plurality of wells of a 96 well plate and measuring 405nM minutes in each well at 6 second intervals using BMG artar galaax plate reader to obtain the rate of catalysis from Graphpad 6.0 and calculate the rate of inhibition in the bolus rate by subtracting the rate of catalytic activity constant (PLOS, see plax constant for rate of catalytic activity, pgas 50).

TABLE 3 EC-based characterization of certain representative compounds₅₀PPI enzyme inhibition of cyclophilin A (CyPA)

CyPA PPI enzyme inhibitory activity: IC₅₀<4.0nM；**4.0nM<IC₅₀<10.0nM；*10.0nM<IC₅₀<20.0nM。

TABLE 4 EC-based characterization of certain representative Compounds₅₀PPI enzyme inhibition of cyclophilin D (CyPD)

CyPA PPI enzyme inhibitory activity: IC₅₀<4.0nM；**4.0nM<IC₅₀<10.0nM；*10.0nM<IC₅₀<20.0nM。

Claims (62)

1. A compound of formula I:

or a pharmaceutically acceptable salt thereof, wherein:

x is 0 or 1;

R₈is alkyl, alkenyl, alkynyl, cycloalkyl, cycloalkenyl, aryl or heteroaryl; wherein R is₈By one or more R₁Substitution; provided that R is₈-R₁Is not n-butyl or (E) -but-2-enyl;

R₂is ethyl, 1-hydroxyethyl, isopropyl or n-propyl;

w is O, S or CH₂；

R₃Is H, alkyl or substituted alkyl, alkenyl or substituted alkenyl, alkynyl or substituted alkynyl, cycloalkyl or substituted cycloalkyl, cycloalkenyl or substituted cycloalkenyl, aryl or substituted aryl or heteroaryl or substituted heteroaryl; wherein R is₃Optionally substituted by one or more R₁Substitution;

R₁independently for each occurrence is H, halogen, aryl OR substituted aryl, heteroaryl OR substituted heteroaryl, CN, OR_A、SR_A、NR_AR_B、-NR_A(CH₂)_OOR_B、-N((CH₂)_OOR_A)((CH₂)_OOR_B)-C(＝O)R_A、-C(＝O)OR_A、-OC(＝O)R_A、-OC(＝O)(C₁-C₆alkyl-R_H)、-OC(＝O)(CH₂)_oOR_A、-C(＝O)NR_AR_B、-NR_AC(＝O)R_B、-N(C(＝O)R_A)(C(＝O)R_B)、-N(C(＝O)(C₁-C₆alkyl-R_H))(C(＝O)(C₁-C₆alkyl-R_H))、-N(C(＝O)(C₁-C₆alkyl-R_H))₂、-NR_AC(＝O)(CH₂)_OOR_B、-NR_AC(＝O)(CH₂)_OOR_B、-N(C(＝O)(CH₂)_OOR_B)₂、-NR_AC(＝O)(CH₂)_ONR_AR_B、-NR_A(CH₂)_OC(＝O)OR_B、-N((CH₂)_OC(＝O)OR_A)((CH₂)_OC(＝O)OR_B)、-NR_A(CH₂)_OC(＝O)NR_AR_B、-N((CH₂)_OC(＝O)NR_AR_B)((CH₂)_OC(＝O)NR_AR_B)、-NR_A(CH₂)_OC(＝O)NR_A(CH₂)_OOR_B、-C(＝O)N((CH₂)_OOR_B)₂、-N((CH₂)_OC(＝O)NR_A(CH₂)_OOR_B)((CH₂)_OC(＝O)NR_A(CH₂)_OOR_B)、-C(＝O)NR_A(CH₂)_OOR_B、-C(＝O)NR_A(CH₂)_OOR_B、-C(＝O)NR_A(CH₂)_ONR_AR_B、-N＝CR_A-NR_AR_B、-NR_B-C(＝NH)-NR_AR_B、O(CH₂)_mOR_A、O(CH₂)_mCOOR_A、O(CH₂)_mCONR_AR_B、O(CH₂)_mCONR_A(CH₂)_mOR_A、O(CH₂)_mO(CH₂)_mOR_A、O(CH₂)_mNR_AR_B、O(CH₂)_mO(CH₂)_mNR_AR_B、NR_C(CH₂)_mNR_AR_B、NR_C(CH₂)_mNR_C(CH₂)_mNR_AR_B、Wherein said aryl or heteroaryl is optionally substituted by a substituent selected from the group consisting of halogen, hydroxy, (C)₁-C₆) Alkyl group, (CH)₂)_pOR_A、(CH₂)_pNR_AR_B、(CH₂)_pC(＝O)NR_AR_BAnd (CH)₂)_pC(＝O)OR_AMay be the same or different, substituted by one or more groups;

R₇is composed of

Each R₅Independently is H, alkyl or substituted alkyl, alkenyl or substituted alkenyl, alkynyl or substituted alkynyl, cycloalkyl or substituted cycloalkyl, cycloalkenyl or substituted cycloalkenyl, or aryl or substituted aryl;

R_Aand R_BIndependently for each occurrence:

hydrogen;

(C₁-C₆) Alkyl, optionally substituted by one or more radicals R, which may be identical or different_DSubstitution;

(C₂-C₆) Alkenyl or (C)₂-C₆) An alkynyl group;

optionally is (C)₁-C₆) Alkyl substituted (C)₃-C₇) A cycloalkyl group;

optionally substituted by a group selected from halogen, -O (C)₁-C₆) Alkyl, -C (═ O) O (C)₁-C₆) Phenyl or benzyl of alkyl, amino, alkylamino and dialkylamino, which may be substituted identically or differently by 1 to 5 radicals;

or may be a saturated or unsaturated heterocyclic ring containing 5 or 6 ring atoms and 1 to 3 heteroatoms, which may be the same or different, selected from nitrogen, sulphur and oxygen;

or R_AAnd R_BTogether with the nitrogen atom to which they are attached form a saturated or unsaturated heterocyclic ring containing from 3 to 7 ring atoms, which may optionally contain another heteroatom selected from nitrogen, oxygen and sulfur and may optionally be substituted by 1 to 4 groups, which may be the same or different, selected from alkyl, phenyl and benzyl;

R_Ceach occurrence is independently hydrogen or (C)₁-C₆) An alkyl group;

R_Deach occurrence is independentGround is halogen, hydroxy, O (C)₁-C₄) Alkyl, C (═ O) (C)₁-C₄) Alkyl, C (═ O) O (C)₁-C₄) Alkyl orWherein Z is CH₂、O、S、NH、NCH₃NEt, N-isopropyl, N-neopentyl, N-CH₂CH₂OH or N-CH₂CH₂OMe；

R_GEach occurrence is independently R_A、OR_A、SR_A、NR_AR_B、–(CH₂)_oR_A、–(CH₂)_oC(＝O)OR_A、–(CH₂)_oC(＝O)NR_AR_B、C(＝O)OR_A、OC(＝O)R_A、NR_AC(＝O)R_B、NR_AC(＝O)(CH₂)_OOR_A、C(＝O)O(CH₂)_OOR_A、C(＝O)OR_B、C(＝O)NR_AR_B、C(＝O)NR_A(CH₂)_OOR_B、C(＝O)N((CH₂)_OOR_A)((CH₂)_OOR_B)、C(＝O)N((CH₂)_OC(＝O)OR_A)((CH₂)_OC(＝O)OR_B)、C(＝O)N((CH₂)_ONR_AR_B)((CH₂)_ONR_AR_B)、C(＝O)N((CH₂)_OOC(＝O)(CH₂)_OOR_A)((CH₂)_OOC(＝O)(CH₂)_OOR_B)、C(＝O)N((CH₂)_ONR_AC(＝O)(CH₂)_OOR_B)((CH₂)_ONR_AC(＝O)(CH₂)_OOR_B)、C(＝O)NR_A(CH₂)_ONR_AR_B、、C(＝O)NR_A(CH₂)_OOC(＝O)R_B、C(＝O)NR_A(CH₂)_OC(＝O)OR_B、C(＝O)NR_A(CH₂)_OC(＝O)NR_AR_B、C(＝O)NR_A(CH₂)_OOC(＝O)(CH₂)_OOR_BOr

R_HIndependently for each occurrence is halogen;

z' is independently CH at each occurrence₂、O、S、NR_A、N(CH₂)_oOR_A、N(CH₂)_oNR_AR_B、N(CH₂)_oCOOR_A、N(CH₂)_oOC(＝O)R_A、N(CH₂)_oCONR_AR_B、N(CH₂)_oNR_AC(＝O)R_BOr N (CH)₂)_oOC(＝O)(CH₂)_oOR_A；

o is independently at each occurrence 0,1, 2,3, 4, 5 or 6;

p is independently at each occurrence an integer of 0,1, 2,3, 4, or 5; and is

m is independently at each occurrence an integer of 1, 2,3, 4 or 5.

2. The compound of claim 1, wherein R₈Is (C)₁-C₁₂) Alkyl, (C)₂-C₁₂) Alkenyl, (C)₂-C₁₂) Alkynyl, (C)₃-C₁₂) Cycloalkyl or phenyl or CH₂-phenyl, optionally substituted by a group selected from halogen, hydroxy, (C)₁-C₆) Alkyl, which may be the same or different, is substituted with one or more groups.

3. A compound according to any one of the preceding claims, wherein R₈Is (C)₁-C₁₄) Alkyl or (C)₁-C₁₄) An alkenyl group.

4. A compound according to any one of the preceding claims, wherein R₈Is (C)₁-C₆) Linear alkyl or (C)₇-C₁₂) A linear alkyl group.

5. A compound according to any one of the preceding claims, wherein R₈Is- (CH)₂)_3-11-an alkyl chain.

6. A compound according to any one of the preceding claims, wherein R₂Is ethyl.

7. A compound according to any one of the preceding claims, wherein R₇Is composed of

8. The compound of any one of the preceding claims, wherein W is O.

9. The compound of any one of the preceding claims, wherein W is S.

10. A compound according to any one of the preceding claims, wherein R₃Is H, (C)₁-C₁₂) Alkyl, (C)₂-C₁₂) Alkenyl, (C)₂-C₁₂) Alkynyl, (C)₃-C₁₂) Cycloalkyl or phenyl or CH₂-phenyl, optionally substituted by a group selected from halogen, hydroxy and (C)₁-C₆) Alkyl, which may be the same or different, is substituted with one or more groups.

11. The compound of claim 1, having the structure of formula (II) or (III):

or a pharmaceutically acceptable salt thereof, wherein:

y is H OR OR₅(ii) a Wherein R is₅Is H or methyl;

m 'and n' are each independently 0,1, 2,3, 4, 5,6, 7,8, 9, 10, 11, or 12;

R_A’and R_B’Independently for each occurrence:

hydrogen;

(C₁-C₆) Alkyl, optionally substituted by one or more radicals R, which may be identical or different_DSubstitution;

(C₂-C₆) Alkenyl or (C)₂-C₆) An alkynyl group;

optionally is (C)₁-C₆) Alkyl substituted (C)₃-C₇) A cycloalkyl group;

optionally substituted by a group selected from halogen, -O (C)₁-C₆) Alkyl, -C (═ O) O (C)₁-C₆) Phenyl or benzyl of alkyl, amino, alkylamino and dialkylamino, which may be substituted identically or differently by 1 to 5 radicals;

or R_A’And R_B’Together with the nitrogen atom to which they are attached form a saturated or unsaturated heterocyclic ring containing from 3 to 7 ring atoms, which may optionally contain another heteroatom selected from nitrogen, oxygen and sulfur and may optionally be substituted by 1 to 4 groups, which may be the same or different, selected from alkyl, phenyl and benzyl; and is

R_DIndependently at each occurrence is halogen, hydroxy, O (C)₁-C₄) Alkyl, C (═ O) (C)₁-C₄) Alkyl, C (═ O) O (C)₁-C₄) Alkyl orWherein Z is CH₂、O、S、NH、NCH₃NEt, N-isopropyl, N-neopentyl, N-CH₂CH₂OH or N-CH₂CH₂OMe。

12. The compound of claim 11, having the structure of formula (IV) or (V):

(ii) a Wherein R is_1’Each occurrence is independently H, halogen, aryl OR substituted aryl, heteroaryl OR substituted heteroaryl, OR_A、SR_A、NR_AR_B、-NR_A(CH₂)_OOR_B、-N((CH₂)_OOR_A)((CH₂)_OOR_B)-C(＝O)R_A、-C(＝O)OR_A、-OC(＝O)R_A、-OC(＝O)(C₁-C₆alkyl-R_H)、-OC(＝O)(CH₂)_oOR_A、-C(＝O)NR_AR_B、-NR_AC(＝O)R_B、-N(C(＝O)R_A)(C(＝O)R_B)、-N(C(＝O)(C₁-C₆alkyl-R_H))(C(＝O)(C₁-C₆alkyl-R_H))、-N(C(＝O)(C₁-C₆alkyl-R_H))₂、-NR_AC(＝O)(CH₂)_OOR_B、-NR_AC(＝O)(CH₂)_OOR_B、-N(C(＝O)(CH₂)_OOR_B)₂、-NR_AC(＝O)(CH₂)_ONR_AR_B、-NR_A(CH₂)_OC(＝O)OR_B、-N((CH₂)_OC(＝O)OR_A)((CH₂)_OC(＝O)OR_B)、-NR_A(CH₂)_OC(＝O)NR_AR_B、-N((CH₂)_OC(＝O)NR_AR_B)((CH₂)_OC(＝O)NR_AR_B)、-NR_A(CH₂)_OC(＝O)NR_A(CH₂)_OOR_B、-C(＝O)N((CH₂)_OOR_B)₂、-N((CH₂)_OC(＝O)NR_A(CH₂)_OOR_B)((CH₂)_OC(＝O)NR_A(CH₂)_OOR_B)、-C(＝O)NR_A(CH₂)_OOR_B、-C(＝O)NR_A(CH₂)_OOR_B、-C(＝O)NR_A(CH₂)_ONR_AR_B、-N＝CR_A-NR_AR_B、-NR_B-C(＝NH)-NR_AR_B、O(CH₂)_mOR_A、O(CH₂)_mCOOR_A、O(CH₂)_mCONR_AR_B、O(CH₂)_mCONR_A(CH₂)_mOR_A、O(CH₂)_mO(CH₂)_mOR_A、O(CH₂)_mNR_AR_B、O(CH₂)_mO(CH₂)_mNR_AR_B、NR_C(CH₂)_mNR_AR_B、NR_C(CH₂)_mNR_C(CH₂)_mNR_AR_B、Wherein said aryl or heteroaryl is optionally substituted by a substituent selected from the group consisting of halogen, hydroxy, (C)₁-C₆) Alkyl group, (CH)₂)_pOR_A、(CH₂)_pNR_AR_B、(CH₂)_pC(＝O)NR_AR_BAnd (CH)₂)_pC(＝O)OR_AMay be the same or different.

13. The compound of claim 12, wherein R_1’Is OR_A、OCOCH₂OR_A、SR_A、NHR_A、N(R_A)₂Or NHCOCH₂OR_A。

14. The compound of claim 12, wherein R_1’Is OAc, OCOCH₂Cl、OCOCH₂CH₃、OCOCHMe₂、OCOCMe₃、OCOCH＝CH₂、NHCH₂CH₂OH、NHCH₂CH₂OMe、N(CH₂CH₂OH)₂、N(CH₂CH₂OMe)₂、NHCH₂CHMe₂、NHCH₂CMe₂、NHAc、NHCH₂COOH、NHCH₂COOCH₃、NMeCH₂COOH、NMeCH₂COOCH₃、NHCH₂CONH₂、NHCH₂CONHMe、NHCH₂CONMe₂、NHCH₂CONHCH₂CH₂OH、NHCH₂CONHCH₂CH₂OMe、NMeCH₂CONH₂、NMeCH₂CONHMe、NMeCH₂CONMe₂、N(CH₂COOH)₂、N(CH₂CONH₂)₂、N(CH₂CONHMe)₂、N(CH₂CONMe₂)₂、N(CH₂CONHCH₂CH₂OH)₂、N(CH₂CONHCH₂CH₂OMe)₂、NHCOCH₂Cl、NHCOCH₂CH₃、NHCOCHMe₂、NHCOCMe₃、NHCOCH＝CH₂、N(COCH₂Cl)₂、N(COCH₂CH₃)₂、N(COCHMe₂)₂、N(COCMe₃)₂Or N (COCH ═ CH)₂)₂。

15. The compound of claim 11 or 12, wherein R_A’And R_B’Each independently of the others is H, Me, Et, n-propyl, isopropyl, isobutyl, neopentyl, cyclopentyl, cyclohexyl, CH₂CH₂OH、CH₂CH₂OMe、Wherein Z is CH₂、O、S、NH、NCH₃NEt, N-isopropyl, N-neopentyl, N-CH₂CH₂OH or N-CH₂CH₂OMe。

16. The compound of claim 12, wherein R₁is-NR_AC(＝O)R_BAnd R is_1’Is OR_A。

17. The compound of claim 16, wherein R₁is-NHC (═ O) R_BAnd R is_1’Is OH.

18. The compound of claim 1, having the structure of formula (VI):

wherein

W is CH₂O or S;

y is H OR OR₅(ii) a Wherein R is₅Is H or methyl;

m 'and n' are each independently 0,1, 2,3, 4, 5,6, 7,8, 9, or 10; and is

R₁Each occurrence is independently H, halogen, aryl OR substituted aryl, heteroaryl OR substituted heteroaryl, OR_A、SR_A、NR_AR_B、-C(＝O)R_A、-C(＝O)OR_A、-C(＝O)NR_AR_B、-NR_AC(＝O)R_B、-NR_AC(＝O)(CH₂)_OR_B、-NR_AC(＝O)(CH₂)_OOR_B、-NR_AC(＝O)(CH₂)_ONR_AR_Bor-C (═ O) NR_A(CH₂)_OR_B。

19. A compound according to any one of the preceding claims, wherein R₁Selected from the group consisting of: H. OR (OR)_A、SR_A、NR_AR_B、-C(＝O)R_A、-C(＝O)OR_A、-C(＝O)NR_AR_B、-NR_AC(＝O)R_B、-NR_AC(＝O)(CH₂)_OR_B、-NR_AC(＝O)(CH₂)_OOR_B、-NR_AC(＝O)(CH₂)_ONR_AR_B、-C(＝O)NR_A(CH₂)_OR_B、-C(＝O)NR_A(CH₂)_OOR_Band-C (═ O) NR_A(CH₂)_ONR_AR_B。

20. A compound according to any one of the preceding claims, wherein R₁Selected from the group consisting of: o (CH)₂)_mOR_A、O(CH₂)_mO(CH₂)_mOR_A、O(CH₂)_mNR_AR_B、O(CH₂)_mO(CH₂)_mNR_AR_B、NR_C(CH₂)_mNR_AR_BAnd NR_C(CH₂)_mNR_C(CH₂)_mNR_AR_B。

21. The method of any preceding claimA compound of formula (I) wherein R₁Selected from the group consisting of: OR (OR)_A、SR_A、NR_AR_B、-C(＝O)R_A、-C(＝O)OR_Aand-C (═ O) NR_AR_B。

22. A compound according to any one of the preceding claims, wherein R₁Is OH, OMe, OEt, O-isopropyl, O-isobutyl, O-neopentyl, O-cyclopentyl, O-cyclohexyl, SH, SMe, S-isopropyl, S-isobutyl, S-neopentyl, S-cyclopentyl, S-cyclohexyl,

23. A compound according to any one of the preceding claims, wherein R₁Is composed ofWherein Z is CH₂、O、S、NH、NCH₃NEt, N-isopropyl, N-neopentyl, N-CH₂CH₂OH or N-CH₂CH₂OMe。

24. A compound according to any one of the preceding claims, wherein R₁Selected from the group consisting of: H. OH, OMe, OEt, O-isopropyl, O-isobutyl, O-neopentyl, O-cyclopentyl, O-cyclohexyl, OCH₂CH₂OH、OCH₂CH₂OCH₃、OCH₂COOH、OCH₂COOCH₃、OCH₂CONH₂、OCH₂CONHMe、OCH₂CONMe₂、OCH₂CONHCH₂CH₂OH、OCH₂CONHCH₂CH₂OMe、OAc、OOCCH₂Cl、OOCCH₂OR_A、OOCCH₂CH₃、OOCCHMe₂、OOCCMe₃And OC (O) CCH (CH)₂。

25. A compound according to any one of the preceding claims, wherein R₁Selected from the group consisting of:

26. a compound according to any one of the preceding claims, wherein R₁Is SH, SMe, SEt, S-isopropyl, S-isobutyl, S-neopentyl, O-cyclopentyl or S-cyclohexyl.

27. A compound according to any one of the preceding claims, wherein R₁Selected from the group consisting of: NH (NH)₂、NHCH₃、NHCH₂CH₃、NHCH₂CHOH、NHCH₂CH₂OMe、NMe₂、NEt₂、NHCH₂CHMe₂、NHCH₂CMe₃、NHAc、NHCH₂COOH、NHCH₂COOCH₃、NMeCH₂COOH、NMeCH₂COOCH₃、NHCH₂CONH₂、NHCH₂CONHMe、NHCH₂CONMe₂、NHCH₂CONHCH₂CH₂OH、NHCH₂CONHCH₂CH₂OMe、NMeCH₂CONH₂、NMeCH₂CONHMe、NMeCH₂CONMe₂、N(CH₂COOH)₂、N(CH₂CONH₂)₂、N(CH₂CONHMe)₂、N(CH₂CONMe₂)₂、N(CH₂CONHCH₂CH₂OH)₂、N(CH₂CONHCH₂CH₂OMe)₂、NHCOCH₂Cl、NHCOCH₂OR_A、NHCOCH₂CH₃、NHCOCHMe₂、NHCOCMe₃、NHCOCH＝CH₂、N(COCH₂Cl)₂、N(COCH₂OR_A)₂、N(COCH₂CH₃)₂、N(COCHMe₂)₂、N(COCMe₃)₂And N (COCH ═ CH)₂)₂。

28. A compound according to any one of the preceding claims, wherein R₁Is composed ofWherein Z is CH₂O, S, NH, NMe, NEt, N-isopropyl, N-neopentyl, N-CH₂CH₂OH、NCH₂CH₂OCH₃、NCH₂CH₂OCH₃、NCH₂COOH、NCH₂COOMe、N-CH₂CONH₂、NCH₂CONHMe or NCH₂CONMe₂。

29. A compound according to any one of the preceding claims, wherein R₁Selected from the group consisting of:

30. a compound according to any one of the preceding claims, wherein R₁Selected from the group consisting of:

wherein Z' is independently at each occurrence CH₂、O、S、NR_A、N(CH₂)_oOR_A、N(CH₂)_oNR_AR_B、N(CH₂)_oCOOR_A、N(CH₂)_oOC(＝O)R_A、N(CH₂)_oCONR_AR_B、N(CH₂)_oNR_AC(＝O)R_BOr N (CH)₂)_oOC(＝O)(CH₂)_oOR_A。

31. A compound according to any one of the preceding claims, wherein R₁is-COOH, -COOMe, -COOEt, -CONH₂、-CONHMe、-CONMe₂、-CONHEt、-CONEt₂、-CONHCH₂CH₂OH、-CONHCH₂CH₂OMe、-CON(CH₂CH₂OH)₂、-CON(CH₂CH₂OMe)₂or-CONMe₂。

32. A compound according to any one of the preceding claims, wherein R₁Selected from the group consisting of:

(ii) a Wherein Z is CH₂、O、S、NH、NCH₃NEt, N-isopropyl, N-neopentyl, N-CH₂CH₂OH or N-CH₂CH₂OMe。

33. A compound according to any one of the preceding claims, wherein R₁Is composed of

34. The compound of claim 33, wherein R_GIs OH, OMe, OAc, NH₂、NHMe、NHAc、NMe₂、NEt₂、NHCH₂CMe₃。

35. A compound according to any one of the preceding claims, wherein R₁Selected from the group consisting of:

36. a compound according to any one of the preceding claims, wherein R₁Is OR_A、OCOCH₂OR_A、SR_A、NHR_A、N(R_A)₂Or NHCOCH₂OR_A。

37. A compound according to any one of the preceding claims, wherein R₁Is OAc, OCOCH₂Cl、OCOCH₂CH₃、OCOCHMe₂、OCOCMe₃、OCOCH＝CH₂、NHCH₂CH₂OH、NHCH₂CH₂OMe、N(CH₂CH₂OH)₂、N(CH₂CH₂OMe)₂、NHCH₂CHMe₂、NHCH₂CMe₂、NHAc、NHCH₂COOH、NHCH₂COOCH₃、NMeCH₂COOH、NMeCH₂COOCH₃、NHCH₂CONH₂、NHCH₂CONHMe、NHCH₂CONMe₂、NHCH₂CONHCH₂CH₂OH、NHCH₂CONHCH₂CH₂OMe、NMeCH₂CONH₂、NMeCH₂CONHMe、NMeCH₂CONMe₂、N(CH₂COOH)₂、N(CH₂CONH₂)₂、N(CH₂CONHMe)₂、N(CH₂CONMe₂)₂、N(CH₂CONHCH₂CH₂OH)₂、N(CH₂CONHCH₂CH₂OMe)₂、NHCOCH₂Cl、NHCOCH₂CH₃、NHCOCHMe₂、NHCOCMe₃、NHCOCH＝CH₂、N(COCH₂Cl)₂、N(COCH₂CH₃)₂、N(COCHMe₂)₂、N(COCMe₃)₂Or N (COCH ═ CH)₂)₂。

38. The compound of claim 1, wherein R₁Is aryl or heteroaryl, said aryl or heteroaryl being optionally substituted by a group selected from halogen, hydroxy, (C)₁-C₆) Alkyl, (C)₃-C₇) Cycloalkyl, SR_A、(CH₂)_pOR_A、(CH₂)_pNR_AR_B、(CH₂)_pC(＝O)R_A、(CH₂)_pC(＝O)NR_AR_BAnd (CH)₂)_pC(＝O)OR_AAnd may be the same or different and is substituted by one or more groups; wherein

p is 0,1, 2,3, 4, 5, 6; and is

R_AAnd R_BIndependently for each occurrence:

hydrogen;

(C₁-C₆) Alkyl, optionally substituted by one or more radicals R, which may be identical or different_DSubstitution;

(C₂-C₆) Alkenyl or (C)₂-C₆) An alkynyl group;

optionally is (C)₁-C₆) Alkyl substituted (C)₃-C₇) A cycloalkyl group;

optionally substituted by a group selected from halogen, -O (C)₁-C₆) Alkyl, -C (═ O) O (C)₁-C₆) Phenyl which is substituted by 1 to 5 groups which may be the same or different, of alkyl, amino, alkylamino and dialkylamino;

or may be a saturated or unsaturated heterocyclic ring containing 5 or 6 ring atoms and 1 to 3 heteroatoms, which may be the same or different, selected from nitrogen, sulphur and oxygen;

or R_AAnd R_BTogether with the nitrogen atom to which they are attached form a saturated or unsaturated heterocyclic ring containing from 3 to 7 ring atoms, which may optionally contain another heteroatom selected from nitrogen, oxygen and sulfur and may optionally be substituted by 1 to 4 groups, which may be the same or different, selected from alkyl, phenyl and benzyl; r_DIndependently at each occurrence is halogen, hydroxy, O (C)₁-C₄) Alkyl, C (═ O) (C)₁-C₄) Alkyl, C (═ O) O (C)₁-C₄) Alkyl orWherein Z is CH₂、O、S、NH、NCH₃NEt, N-isopropyl, N-neopentyl, N-CH₂CH₂OH or N-CH₂CH₂OMe。

39. The compound of claim 1, wherein R₁Is composed ofWherein Rx is H, (C)₁-C₆) Alkyl or (C)₃-C₇) A cycloalkyl group; ry is H, (C)₁-C₆) Alkyl, (C)₃-C₇) Cycloalkyl, OR_A、SR_A、NR_AR_B、-C(＝O)R_A、-C(＝O)OR_Aor-C (═ O) NR_AR_B(ii) a And t is 1, 2,3 or 4.

40. The compound of claim 39, wherein Rx is H or Me; and Ry is-C (═ O) OR_Aor-C (═ O) NR_AR_B。

41. A compound according to claim 40, wherein R₁Is composed ofWherein Z is CH₂、O、S、NH、NCH₃NEt, N-isopropyl, N-neopentyl, N-CH₂CH₂OH or N-CH₂CH₂OMe。

42. A compound according to any one of the preceding claims, wherein R_AAnd R_BEach independently of the others is H, Me, Et, n-propyl, isopropyl, isobutyl, neopentyl, cyclopentyl, cyclohexyl, CH₂CH₂OH、CH₂CH₂OMe、Wherein Z is CH₂、O、S、NH、NCH₃NEt, N-isopropyl, N-neopentyl, N-CH₂CH₂OH or N-CH₂CH₂OMe。

43. A compound according to claim 42, wherein R_AIs H, Me, Et, n-propyl, isopropyl, n-butyl or isobutyl.

44. A compound according to claim 42, wherein R_AAnd R_BEach independently is H, Me, Et, isopropyl, isobutyl, cyclopentyl, or cyclohexyl.

45. A compound according to any one of the preceding claims, wherein R_AAnd R_BIndependently at each occurrence, H, (C)₁-C₆) Alkyl, phenyl, CH₂-phenyl, (C)₁-C₆) alkyl-OH, (CH)₂)_pO(CH₂)_mOH or (CH)₂)_pO(CH₂)_mO(CH₂)_mOH、(C₁-C₆) alkyl-O- (C)₁-C₄) Alkyl group, (CH)₂)_pO(CH₂)_mO(C₁-C₄) Alkyl or (CH)₂)_pO(CH₂)_mO(CH₂)_mO(C₁-C₄) An alkyl group.

46. A compound according to any one of the preceding claims, wherein R_AAnd R_BEach occurrence is independently H or (C)₁-C₆) An alkyl group.

47. A compound according to claim 46, wherein R_AAnd R_BEach independently is H, Me, Et, isopropyl, isobutyl, cyclopentyl, or cyclohexyl.

48. A compound according to any one of the preceding claims, wherein R_AAnd R_BTogether with the nitrogen atom to which they are attachedIs selected fromWherein R is_CIs H, Me, Et, n-Pr, i-Pr, n-Bu, i-Bu, t-Bu, CH₂CMe₃、Ph、CH₂Ph、CH₂CH₂OH or CH₂CH₂O(C₁-C₄) An alkyl group.

49. The compound of claim 1, having a structure selected from the group consisting of:

wherein x is 0 or 1; m is 2,3, 4, 5,6, 7 or 8; y is H, OH or OMe; w is O or S; and each R_aIndependently selected from the group consisting of the moieties listed in table 1:

TABLE 1

，

Or a pharmaceutically acceptable salt thereof.

50. The compound of any one of claims 1-49, wherein x is 0.

51. The compound of any one of claims 1-49, wherein x is 1.

52. A pharmaceutical composition comprising at least one compound according to any one of claims 1-51 and a pharmaceutically acceptable carrier or diluent.

53. A method for treating or preventing a viral infection in a mammalian species in need thereof, the method comprising administering to the mammalian species a therapeutically effective amount of at least one compound of any one of claims 1-51.

54. A method for treating or preventing a hepatitis C virus infection in a mammalian species in need thereof, the method comprising administering to the mammalian species a therapeutically effective amount of at least one compound of any one of claims 1-51.

55. A method for treating or preventing a hepatitis B virus infection in a mammalian species in need thereof, the method comprising administering to the mammalian species a therapeutically effective amount of at least one compound of any one of claims 1-51.

56. A method for treating or preventing HIV infection in a mammalian species in need thereof, the method comprising administering to the mammalian species a therapeutically effective amount of at least one compound of any one of claims 1-51.

57. A method for inhibiting cyclophilin in a subject in need thereof, comprising administering to the subject an amount of at least one compound according to any one of claims 1-51 effective to inhibit cyclophilin.

58. A method for treating or preventing a cyclophilin-mediated disease in a mammalian species in need thereof, comprising administering to the mammalian species a therapeutically effective amount of at least one compound of any one of claims 1-51.

59. A method for treating or preventing a disease in a mammalian species in need thereof, the method comprising administering to the mammalian species a therapeutically effective amount of at least one compound of any one of claims 1-51, wherein the disease is selected from inflammation, respiratory inflammation, rheumatoid arthritis, and dry eye.

60. A method for treating or preventing a disease in a mammalian species in need thereof, the method comprising administering to the mammalian species a therapeutically effective amount of at least one compound of any one of claims 1-51, wherein the disease is selected from the group consisting of neurodegenerative diseases such as Alzheimer's disease, Parkinson's disease, Huntington's disease, and ALS; traumatic brain injury; stroke; ischemia reperfusion injury in brain, heart, kidney, and myocardial infarction.

61. A method for the treatment or prevention of a disease in a mammalian species in need thereof, the method comprising administering to the mammalian species a therapeutically effective amount of at least one compound according to any one of claims 1-51, wherein the disease is selected from cardiovascular disease, vascular stenosis, atherosclerosis, abdominal aortic aneurysm, cardiac hypertrophy, aortic rupture, pulmonary hypertension, myocarditis, and myocardial fibrosis and ischemic heart disease.

62. A method for treating or preventing a disease or condition in a mammalian species in need thereof, the method comprising administering to the mammalian species a therapeutically effective amount of at least one compound of any one of claims 1-51, wherein the disease or condition is selected from the group consisting of cancer, obesity, diabetes, muscle atrophy, lung disease, liver disease, kidney disease, and hair loss.