HK1188134A

HK1188134A - Novel cyclosporin derivatives for the treatment and prevention of a viral infection

Info

Publication number: HK1188134A
Application number: HK14101275.8A
Authority: HK
Inventors: 苏壮; 龙正宇; 黄震年; 杨遂周
Original assignee: 河北鲲翔济世医药科技有限公司
Priority date: 2010-12-03
Filing date: 2011-12-05
Publication date: 2014-04-25

Abstract

The present invention relates to a compound of the Formula (I): or pharmaceutically acceptable salt thereof, wherein the symbols are as defined in the specification; a pharmaceutical composition comprising the same, a method for treating or preventing a viral infection using the same.

Description

Novel cyclosporin derivatives for the treatment and prevention of viral infections

Cross Reference to Related Applications

This application claims the benefit of U.S. provisional application No. 61/419,326, filed on 3/12/2010, the entire contents of which are incorporated herein by reference.

Technical Field

The present invention relates to novel cyclosporin derivatives, pharmaceutical compositions containing the same and methods of using the same for treating or preventing viral infections.

Background

Naturally occurring cyclosporines are poly-N-methyl, cyclic undecapeptides isolated from fungi. Cyclosporin a has immunosuppressive activity and has been used to prevent rejection in kidney, heart and liver transplant recipients for nearly 40 years. It has anti-inflammatory properties and is useful for the treatment of rheumatoid arthritis, severe psoriasis, Behget uveitis and dry eye. In addition, it is useful for the treatment of severe ulcerative colitis, Crohn's disease, alopecia areata, aplastic anemia, HSV-1 keratitis, systemic lupus erythematosus and severe lupus nephritis.

Cyclosporin a was found to have anti-HIV activity (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, 1067-1078; each of which is incorporated herein by reference). Its non-immunosuppressive derivative NIM-811 is reported to have potent anti-HIV activity due to its ability to inhibit cyclophilin A (Franke, E.K., et al, 1994, Nature, 372, 359-362; Thali, M., et al, 1994, Nature, 372, 363-365; Gamble, T.R., et al, 1996, Cell, 87, 1157-1159; Rosenwirth B., et al, 1994, Antimicrob. Agents Chemother., 38, 1763-1772; 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 and SCY-635, inhibit cyclophilin, which interacts with HCV protein NS5B and stimulates the RNA-binding activity of NS 5B. Thus, 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.gastroentenol., 38, 567-72; Watashi, k., et al, 2003, Hepatology, 38, 1282-8; gaitherer, l.a., et al, 2010, Virology, 397, 43-55; each of which is incorporated herein by reference). At present, NIM-811, Debio-025 and SCY-635 are undergoing clinical trials for the treatment of HCV.

NIM-811 and Debio-025 have a chemical structure similar to cyclosporin a and have poor pharmacokinetic profiles and poor oral absorption. In addition, they are metabolized by P450 to induce drug interactions (Lill, J., et al, 2000, Am J Health-Syst Pharm 57, 1579; incorporated herein by reference).

SCY-635 has an improved pharmacokinetic profile and low serum binding. In addition, it is less metabolized by P450 and has a low potential for drug-drug interactions. In vitro anti-HCV Activity of SCY-635 (EC) by the luciferase endpoint method developed by using Hopkins, S. et al, 2010, Antimicrob.Agents Chemother, 54, 660-672, incorporated herein by reference₅₀) Reported as 0.10 μ M. However, SCY-635 is chemically unstable according to the test results of our laboratory. SCY-635 is readily converted to its diastereomer by epimerization, which is expected to have poor binding activity for cyclophilin and thus may affect its antiviral activity in vivo.

Cyclosporin a and its non-immunosuppressive derivatives were also found to possess anti-HBV activity by inhibiting cyclophilin (Chokshi, s., et al, 2011, Abstract 190 (chest Presentations), 46th annual Meeting of the European Association for the Study of the Liver (EASL2011), Berlin, March 30-April 3; Tian, x.c., et al, 2010, j.virol., 84, 3373-3381; Xia, w.l., et al, 2004, hepatobiliar Pancreat disease int, 4, 18-22; Michael, j., et al, 2003, j.virol., 77, 7713-7719; each of which is incorporated herein by reference).

In addition, cyclophilins are reported to regulate the life cycle and pathogenesis of a variety of viruses including influenza a viruses, severe acute respiratory syndrome coronaviruses, and vaccinia viruses (Castro, a.p., et al, 2003, j.virol., 77, 9052-9068; Chen, z., l., et al, 2005, j.infact.dis.191, 755-760; Liu, x.l., et al, 2009, microcell biol.11, 730-741; each of which is incorporated herein by reference). Cyclosporin a and its non-immunosuppressive derivatives also possess such antiviral activity.

N-MeVal-4-cyclosporin (SDZ 220-384), another non-immunosuppressive cyclosporin derivative, has been reported to have biological activity similar to NIM-811 (Fliri, H., et al, 1993, Ann.N YAcad 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 of the Flaviviridae family. HCV has a high replication rate and an abnormally high mutation rate. Most people infected with HCV (about 80%) develop chronic, persistent infections. Over 400 million americans have been infected with HCV and over 2 million people worldwide are expected to have chronic infections. Approximately 35,000 new cases of hepatitis c are expected to occur in the united states annually. In the united states, HCV infection causes about 50% of all chronic liver diseases, 30% of all liver transplants, and 30% of all liver cirrhosis, end stage liver diseases, and liver cancers. peg-interferon and ribavirin combinations are standard treatments for chronic hepatitis c but have low efficacy against HCV infection. Recently, the FDA approved litex's inclik (telaprevir) and Merck's Victrelis (boceprevir) as adjunctive therapies to current interferon/ribavirin therapies for the treatment of HCV. Both drugs are HCV protease inhibitors and target the virus to prevent its replication. However, due to the rapid mutation of HCV, drug resistance to new drugs can be developed in a short time. There is a need for effective therapeutic agents for the treatment of HCV.

Hepatitis B Virus (HBV) is a 42nm partially double-stranded DNA virus consisting essentially of a 27nm core-coat protein core (HBcAg) surrounded by an outer lipoprotein envelope containing surface antigens (HBsAg). About one quarter of the world's population, over 20 million people, has been infected with hepatitis b virus. This includes long-term carriers of 3.50 billion viruses. The disease causes epidemics in parts of asia and africa, and it is endemic in china. Chronic hepatitis b will lead to cirrhosis and liver cancer, a fatal disease that responds very poorly to current chemotherapy. Although this infection is preventable by vaccination and HBV load and replication can be reduced by the current antiviral drugs lamivudine (Epivir), adefovir (hepera), tenofovir (Viread), telbivudine (Tyzeka) and entecavir (Baraclude) and the two immune system modulators interferon alpha-2 a and pegylated interferon alpha-2 a (Pegasys), none of the available drugs is able to clear the infection. There remains a need for effective therapeutic agents for treating or preventing HBV infection.

Non-immunosuppressive cyclosporin derivatives bind to cyclophilins, a family of host proteins that catalyze cis-trans peptidyl-prolyl isomerization in the protein folding critical for processing, maturation of viral proteins for viral replication. It also differs from current anti-HIV and anti-HCV drugs, the advantage of cyclosporin derivatives targeting host cofactor-cyclophilin is a presumed higher genetic Barrier to resistance to development (Rosenwirth, b., et al, 1994, anti. ingredients chemi., 38, 1763-1772; Tang, h.l. et al, 2010, virues, 2, 1621-1634; Hopkins, s. et al, 2010, Oral Presentation, Scynexis's SCY-635 expressed Barrier to HCV Treatment, the 45th Annual Meeting of the european association for the Study of the 2010 Liver (EASL), Vienna, stria, April 14-18; each of which is incorporated herein by reference). Cyclosporin derivatives affect a new target, cyclophilin, and thus represent a new mechanism of action against HCV.

Cyclophilins are a family of enzymes that aid in the folding and transport of other proteins synthesized in cells. Protein folding or misfolding plays an important role in the pathophysiology of many serious diseases such as: viral diseases (HCV, HIV, and herpes simplex virus), central nervous system disorders (stroke, traumatic brain and spinal injuries, alzheimer's disease, parkinson's disease, and mitochondrial protection of huntington's disease), cardiovascular diseases (myocardial infarction, heart attack, and reperfusion injury in chronic heart failure), cancer, inflammation (respiratory inflammation, asthma, rheumatoid arthritis, dry eye disease) and including inflammatory bowel disease (crohn's disease and ulcerative colitis), atopic dermatitis, antifungal and antiparasitic treatments, hair growth, and obesity, diabetes, muscular dystrophy, and NSAID-induced bowel disease.

Cyclosporin derivatives target cyclophilins and may play a key role in the treatment of these diseases.

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, 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.

Summary of The Invention

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

or a pharmaceutically acceptable salt thereof, wherein:

R₈is n-butyl, (E) -but-2-enyl or

R₂Is ethyl, 1-hydroxyethyl, isopropyl or n-propyl;

w is O, S or NR₁；

R₁Is 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 heterocyclic or substituted heterocyclic; or R₁And R₃Together with the nitrogen atom to which they are attached form a saturated or unsaturated heterocyclic ring containing three to seven ring atoms, which ring may optionally contain another heteroatom selected from the group consisting of nitrogen, oxygen and sulfur, and may optionally be substituted by one to four groups selected from the group consisting of alkyl, phenyl and benzyl, which may be the same or different;

R₃is 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;

R₇is thatAnd is

Each R₅Independently is H, alkyl or substituted alkyl, alkenyl or substituted alkeneA group, alkynyl or substituted alkynyl, cycloalkyl or substituted cycloalkyl, cycloalkenyl or substituted cycloalkenyl or aryl or substituted aryl.

In another aspect, the present invention provides a compound of formula (I) as shown above, or a pharmaceutically acceptable salt thereof, wherein:

R₈is n-butyl, (E) -but-2-enyl or

R₂Is ethyl, 1-hydroxyethyl, isopropyl or n-propyl;

w is O, S or NR₁；

R₁Is 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;

(C₃-C₇) Cycloalkyl, optionally with (C)₁-C₆) Alkyl substitution;

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

or a heterocyclic ring, which may be saturated or unsaturated, containing five or six ring atoms and one to three heteroatoms, which may be the same or different, selected from nitrogen, sulfur and oxygen;

or R₁And R₃Together with the nitrogen atom to which they are attached form a saturated or unsaturated heterocyclic ring containing three to seven ring atoms, which may optionally contain another heteroatom selected from the group consisting of nitrogen, oxygen and sulfur, and mayOptionally selected from (C), which may be the same or different₁-C₆) One to four groups of the group consisting of alkyl, phenyl and benzyl;

R₃the method comprises the following steps:

H；

(C₁-C₆) Alkyl, optionally substituted by one or more radicals R which may be the same or different₄Substitution;

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

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

(C₃-C₇) Cycloalkyl optionally substituted by one or more groups which may be the same or different selected from halo, hydroxy, amino, monoalkylamino and dialkylamino;

phenyl or CH₂-phenyl, optionally substituted by a group which may be the same or different selected from halogen, hydroxy, (C)₁-C₆) One or more groups of alkyl are substituted;

R₇is that

R₅The method comprises the following steps:

H；

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

(C₂-C₆) Alkenyl, optionally substituted by a group which may be the same or different selected from 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);

phenyl or CH₂-phenyl, optionally substituted by a group which may be the same or different selected 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);

each occurrence of R₄Independently halogen, hydroxy, aryl (e.g. phenyl), O (CH)₂)_mOR_A、O(CH₂)_mO(CH₂)_mOR_A、C(＝O)(C₁-C₆) Alkyl, C (═ O) OR_A、C(＝O)NR_AR_B、-NR_AR_B、-NR_CCH₂(CH₂)_pNR_AR_B、NR_C[CH₂(CH₂)_pNR_A]_mCH₂(CH₂)_nNR_AR_B、O[CH₂(CH₂)_pO]_mCH₂(CH₂)_nOR_A、OCH₂(CH₂)_pNR_AR_BOr O [ CH ]₂(CH₂)_pO]_mCH₂(CH₂)_nNR_AR_B；

Each occurrence of R₆Independently 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 by a group which may be the same or different 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 occurrence of R_AAnd R_BIndependently are:

hydrogen;

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

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

(C₃-C₇) Cycloalkyl, optionally with (C)₁-C₆) Alkyl substitution;

or R_AAnd R_BTogether with the nitrogen atom to which they are attached, form a saturated or unsaturated heterocyclic ring containing three to seven ring atoms, which ring may optionally contain another heteroatom selected from the group consisting of nitrogen, oxygen and sulfur, and may optionally be substituted by one to four groups which may be the same or different selected from the group consisting of alkyl, phenyl and benzyl;

each occurrence of R_CIndependently is hydrogen or (C)₁-C₆) An alkyl group;

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

m is an integer of 1, 2, 3, 4 or 5.

In another aspect, the present invention provides compounds of formulae (II) to (V):

or a pharmaceutically acceptable salt thereof, wherein:

represents a single or double bond;

each W is independently O, S or NR₁；

Each R₁Independently is 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 a heterocyclic or substituted heterocyclic ring; or R₁And R₃Together with the nitrogen atom to which they are attached form a saturated or unsaturated heterocyclic ring containing three to seven ring atoms, which ring may optionally contain another heteroatom selected from the group consisting of nitrogen, oxygen and sulfur, and may optionally be substituted by one to four groups selected from the group consisting of alkyl, phenyl and benzyl, which may be the same or different; and is

Each occurrence of R₃And 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.

In another aspect, the present invention provides a compound of formulae (II) to (V) as shown above, or a pharmaceutically acceptable salt thereof, wherein:

each W is independently O, S or NR₁；

Each R₁Independently is hydrogen;

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

(C₃-C₇) Cycloalkyl, optionally with (C)₁-C₆) Alkyl substitution;

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

or R₁And R₃Together with the nitrogen atom to which they are attached form a saturated or unsaturated heterocyclic ring containing three to seven ring atoms, which ring may optionally contain another heteroatom selected from the group consisting of nitrogen, oxygen and sulfur, and may optionally be selected from (C) which may be the same or different₁-C₆) One to four groups of the group consisting of alkyl, phenyl and benzyl;

each R₃Independently are:

H；

(C₂-C₆) Alkenyl, optionally substituted by one or more substituents which may be the same or different, 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);

each R₅Independently are:

H；

each occurrence of R_AAnd R_BIndependently are:

hydrogen;

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

(C₃-C₇) Cycloalkyl, optionally with (C)₁-C₆) Alkyl substitution;

phenyl, optionally substituted by a group which may be the same or different selected from halogen, -O (C)₁-C₆) Alkyl, -C (＝O)O(C₁-C₆) One to five groups of alkyl, amino, alkylamino and dialkylamino;

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’；

Each occurrence of R_CIndependently is hydrogen or (C)₁-C₆) An alkyl group;

each occurrence of R_DIndependently halogen, hydroxy, O (C)₁-C₄) Alkyl, C (═ O) (C)₁-C₄) Alkyl, C (═ O) O (C)₁-C₄) An alkyl group;

each occurrence of R_FAnd R_F’Independently of each other, 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 three to seven ring atoms, which ring may optionally contain another heteroatom selected from the group consisting of nitrogen, oxygen and sulfur, and may optionally be substituted by one to four groups which may be the same or different selected from the group consisting of alkyl, phenyl and benzyl;

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

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

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

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

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

Detailed Description

Definition of

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

The terms "alkyl" and "alkyl" refer to a straight or branched alkane (hydrocarbon) group containing 1 to 12 carbon atoms, preferably 1 to 6 carbon atoms. Exemplary "alkyl" groups include methyl, ethyl, propyl, isopropyl, n-butyl, t-butyl, isobutyl, pentyl, hexyl, isohexyl, heptyl, 4-dimethylpentyl, octyl, 2, 4-trimethylpentyl, nonyl, decyl, undecyl, dodecane, and the like. Term "(C)₁-C₄) Alkyl "means a straight or branched chain alkane (hydrocarbon) group containing 1 to 4 carbon atoms, such as methyl, ethyl, propyl, isopropyl, n-butyl, tert-butyl and isobutyl. Term "(C)₁-C₆) Alkyl means containing 1 to 6Straight-chain or branched-chain alkane (hydrocarbon) groups of carbon atoms, such as n-hexyl, 2-methylpentyl, 3-methylpentyl, 4-methylpentyl, 2, 3-dimethylbutyl, 2-dimethylbutyl, and p- (C)₁-C₄) Alkyl groups "are exemplary. "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 carrying Cl₃Alkyl group of (i), cyano, nitro, oxygen (i.e., ═ O), CF), and the like₃、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 present at each occurrence_aIndependently is hydrogen, alkyl, cycloalkyl, alkenyl, cycloalkenyl, alkynyl, heterocycle, or aryl; each occurrence of R_b、R_cAnd R_dIndependently is 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 at each occurrence_eIndependently is alkyl, cycloalkyl, alkenyl, cycloalkenyl, alkynyl, heterocycle or aryl. In thatOf the foregoing 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-chain or branched hydrocarbon group having 2 to 6 carbon atoms and at least one carbon-carbon double bond, such as vinyl, 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, m-n-2, (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 an alkenyl 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 carrying Cl₃Alkyl group of (i), cyano, nitro, oxygen (i.e., ═ O), CF), and the like₃OCF3, cycloalkyl, 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 present at each occurrence_aIndependently is hydrogen, alkyl, cycloalkyl, alkenyl, cycloalkenyl, alkynyl, heterocycle, or aryl; each occurrence of R_b、R_cAnd R_dIndependently is 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 at each occurrence_eIndependently is alkyl, cycloalkyl, alkenyl, cycloalkenyl, alkynyl, heterocycle or aryl. Exemplary substituents may themselves be optionally substituted.

The term "alkynyl" refers to a straight or branched chain hydrocarbon group containing 2 to 12 carbon atoms and at least one carbon-carbon triple bond. Examples of such groups include ethynyl. The term "C₂-C₆Alkynyl "refers to 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 an alkynyl 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 carrying Cl₃Alkyl group of (i), cyano, nitro, oxygen (i.e., ═ O), CF), and the like₃OCF3, cycloalkyl, 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 present at each occurrence_aIndependently is hydrogen, alkyl, cycloalkyl, alkenyl, cycloalkenyl, alkynyl, heterocycle, or aryl; each occurrence of R_b、R_cAnd R_dIndependently is 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 at each occurrence_eIndependently is alkyl, cycloalkyl, alkenyl, cycloalkenyl, alkynyl, heterocycle or aryl. Exemplary substituents may themselves be optionally substituted.

The term "cycloalkyl" refers to a fully saturated cyclic hydrocarbon group containing 1 to 4 rings and 3 to 8 carbons 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 carrying Cl₃Alkyl group of (i), cyano, nitro, oxygen (i.e., ═ O), CF), and the like₃OCF3, cycloalkyl, 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 present at each occurrence_aIndependently is hydrogen, alkyl, cycloalkyl, alkenyl, cycloalkenyl, alkynyl, heterocycle, or aryl; each occurrence of R_b、R_cAnd R_dIndependently is 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 at each occurrence_eIndependently is alkyl, cycloalkyl, alkenyl, cycloalkenyl, alkynyl, heterocycle or aryl. Exemplary substituents may themselves be optionally substituted. Exemplary substituents also include spiro-linked or fused-ring substituents, especially spiro-linked cycloalkyls, spiro-linked cycloalkenyls, spiro-linked heterocycles (excluding heteroaryl), fused-ring alkyls, fused-ring alkenyls, fused heterocycles, or fused aryl groups, wherein the aforementioned cycloalkyl, cycloalkenyl, heterocycle, and aryl substituents can themselves be optionally substituted.

The term "cycloalkenyl" refers to a partially unsaturated cyclic hydrocarbon group containing 1 to 4 rings and 3 to 8 carbons per ring. Examples of such groups include cyclobutenyl, cyclopentenyl, cyclohexenyl, and the like. "substituted cycloalkenyl" refers to a cycloalkenyl 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 carrying Cl₃Alkyl group of (i), cyano, nitro, oxygen (i.e., ═ O), CF), and the like₃、OCF₃Cycloalkyl, alkenyl, cycloalkenyl, alkyneRadical, heterocyclic, 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 present at each occurrence_aIndependently is hydrogen, alkyl, cycloalkyl, alkenyl, cycloalkenyl, alkynyl, heterocycle, or aryl; each occurrence of R_b、R_cAnd R_dIndependently is 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 at each occurrence_eIndependently is alkyl, cycloalkyl, alkenyl, cycloalkenyl, alkynyl, heterocycle or aryl. Exemplary substituents may themselves be optionally substituted. Exemplary substituents also include spiro-linked or fused-ring substituents, especially spiro-linked cycloalkyls, spiro-linked cycloalkenyls, spiro-linked heterocycles (excluding heteroaryl), fused-ring alkyls, fused-ring alkenyls, fused heterocycles, or fused aryl groups, wherein the aforementioned cycloalkyl, cycloalkenyl, heterocycle, and aryl substituents can themselves 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.), the aromatic rings of the aryl group can be linked at a single point (e.g., biphenyl) or fused (e.g., naphthyl, phenanthrenyl)(phenylanthrenyl) and the like). "substituted aryl" refers to an aryl group substituted at any available point of attachment with one or more substituents, preferably 1 to 3 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 carrying Cl₃Alkyl group of (i), cyano, nitro, oxygen (i.e., ═ O), CF), and the like₃OCF3, cycloalkyl, 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 present at each occurrence_aIndependently is hydrogen, alkyl, cycloalkyl, alkenyl, cycloalkenyl, alkynyl, heterocycle, or aryl; each occurrence of R_b、R_cAnd R_dIndependently is 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 at each occurrence_eIndependently is alkyl, cycloalkyl, alkenyl, cycloalkenyl, alkynyl, heterocycle or aryl. Exemplary substituents may themselves be optionally substituted. Exemplary substituents also include fused ring groups, especially fused ring alkyl, fused ring alkenyl, fused heterocycle, or fused aryl, wherein the aforementioned cycloalkyl, cycloalkenyl, heterocycle, and aryl groups are taken to beThe substituent may itself be optionally substituted.

The terms "heterocycle" and "heterocyclic" refer to a fully saturated or partially or fully unsaturated 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, including aromatic cyclic groups (i.e., "heteroaryl"). 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 "heteroarylium" refers to a heteroaryl group that carries a quaternary ammonium nitrogen atom and thus a positive charge.) the heterocyclic group may be attached to the remainder of the molecule at any heteroatom or carbon atom of a ring or ring system. Exemplary monocyclic heterocyclic groups include azetidinyl, pyrrolidinyl, pyrrolyl, pyrazolyl, glycidyloxy, pyrazolinyl, imidazolyl, imidazolinyl, imidazolidinyl, oxazolyl, oxazolidinyl, isoxazolinyl, isoxazolyl, thiazolyl, thiadiazolyl, thiazolidinyl, isothiazolyl, isothiazolidinyl, furyl, tetrahydrofuryl, thienyl, oxadiazolyl, piperidinyl, piperazinyl, 2-oxopiperazinyl, 2-oxopiperidinyl, 2-oxopyrrolidinyl, 2-oxoazenyl, azepinyl, hexahydrodiazanyl, 4-piperidinonyl, pyridinyl, pyrazinyl, pyrimidinyl, pyridazinyl, triazinyl, triazolyl, tetrazolyl, tetrahydropyranyl, morpholinyl, thiomorpholinyl sulfoxide, thiomorpholinyl sulfone, 1, 3-dioxolane and tetrahydro-1, 1-dioxythienyl and the like. Exemplary bicyclic heterocyclic groups include indolyl, isoindolyl, benzothiazolyl, benzoxazolyl, benzooxadiazolyl, benzothienyl, benzo [ d ] [1, 3] dioxolanyl, 2, 3-dihydrobenzo [ b ] [1, 4] dioxinyl, quinuclidinyl, quinolinyl, tetrahydroisoquinolinyl, isoquinolinyl, benzimidazolyl, benzopyranyl, indolizinyl, benzofuranyl, benzofurazanyl, chromonyl, coumarinyl, benzopyranyl, cinnolinyl, quinoxalinyl, indazolyl, pyrrolopyridyl, furopyridyl (such as furo [2, 3-c ] pyridyl, furo [3, 2-b ] pyridyl ] or furo [2, 3-b ] pyridyl), dihydroisoindolyl, dihydroquinazolinyl (such as 3, 4-dihydro-4-oxo-quinazolinyl), Triazinylazepinyl, tetrahydroquinolinyl, and the like. Exemplary tricyclic heterocyclic groups include carbazolyl, benzidolyl, phenanthrolinyl, acridinyl, phenanthridinyl, xanthenyl and the like.

"substituted heterocycle" and "substituted heterocycle" (such as "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 carrying Cl₃Alkyl group of (i), cyano, nitro, oxygen (i.e., ═ O), CF), and the like₃、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 present at each occurrence_aIndependently is hydrogen, alkyl, cycloalkyl, alkenyl, cycloalkenyl, alkynyl, heterocycle, or aryl; each occurrence of R_b、R_cAnd R_dIndependently 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 at each occurrence_eIndependently is alkyl, cycloalkyl, alkenyl, cycloalkenyl, alkynyl, heterocycle or aryl. Exemplary substituents may themselves be optionally substituted. Exemplary substituents also include spiro-linked or fused-ring substituents at any available point or points of attachment, especially spiro-linked cycloalkyls, spiro-linked cycloalkenyls, spiro-linked heterocycles (excluding heteroaryl), fused cycloalkyl groups, fused alkenyl groups, fused heterocycles, or fused aryl groups, where the aforementioned cycloalkyl, cycloalkenyl, heterocycle, and aryl substituents can themselves 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 groups include, but are not limited to, methylamino, ethylamino, n-propylamino, i-propylamino, cyclopropylamino, n-butylamino, t-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. In the dialkylamino moiety, R and R' can be the same or different. Examples of dialkylamino groups 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 ring structure. The resulting ring 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-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 unsatisfied 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 include reference to a salt thereof. As used herein, the term "salt" refers to an acidic and/or basic salt formed with an inorganic and/or organic acid and a base. In addition, when the compounds of the present invention contain 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 "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 present invention may be formed, for example, by: compound I is reacted with an amount of, e.g., an equivalent amount of an acid or base in a medium such as one in which a salt precipitates or in an aqueous medium followed by lyophilization.

Compounds of the invention containing a basic moiety such as, but not limited to, an amine or pyridine or imidazole ring may form salts with a variety of organic and inorganic acids. Exemplary acid addition salts include acetates (such as those formed with acetic acid or trihaloacetic acid, e.g., trifluoroacetic acid), adipates, alginates, ascorbates, aspartates, benzoates, benzenesulfonates, bisulfates, borates, butyrates, citrates, camphorates, camphorsulfonates, cyclopentanepropionates, digluconates, dodecylsulfonates, ethanesulfonates, fumarates, glucoheptanoates, glycerophosphates, hemisulfates, heptanoates, caproates, hydrochlorides, hydrobromides, hydroiodides, hydroxyethanesulfonates (e.g., 2-hydroxyethanesulfonates), lactates, maleates, methanesulfonates, naphthalenesulfonates (e.g., 2-naphthalenesulfonates), nicotinates, nitrates, oxalates, pectinates, persulfates, phenylpropionates (e.g., 3-phenylpropionates), Phosphates, picrates, pivalates, propionates, salicylates, succinates, sulfates (such as those formed with sulfuric acid), sulfonates, tartrates, thiocyanates, tosylates (tolumene-4-sulfonate) such as tosylate (tosilate), undecanoates, and the like.

The compounds of the present invention containing an acidic moiety such as, but not limited to, carboxylic acids 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 benzathines, dicyclohexylamines, hydrabamines (formed with N, N-bis (dehydroabietyl) ethylenediamine), N-methyl-D-glucamines, N-methyl-D-bisamides, t-butyl amines, 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 employed herein refers to a compound that undergoes chemical conversion by metabolic or chemical processes to yield a compound of the invention, or a salt and/or solvate thereof, when administered 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 (e.g., 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 carbons on different substituents), including enantiomeric forms and diastereomeric forms, are contemplated within the scope of the invention. The various 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 the indicated activity), or may be mixed, for example, as a racemic mixture or as a mixture with all other stereoisomers or other selected stereoisomers. The chiral centers of the present invention may have the S or R configuration as defined by International Union of Pure and Applied Chemistry (IUPAC)1974 Recommendations. 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. Individual optical isomers may be obtained from the racemic mixture by any suitable method, including, without limitation, conventional methods such as, for example, salt formation with an optically active acid followed by crystallization.

The compounds of the present invention are preferably isolated and purified after their preparation to obtain a composition containing an amount equal to or greater than 90%, e.g., equal to or greater than 95%, equal to or greater than 99% pure ("substantially pure" compound I) by weight, 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, whether in admixture or 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, groups and substituents thereof may be selected to provide stable moieties and compounds.

The definitions of particular functional groups and chemical terms are described in more detail below. For the purposes of the present invention, chemistryElements are identified according to the following: the Periodic Table of the Elements, CAS version, Handbook of Chemistry and Physics, 75^thEd, cover sheet, and the specific functional groups are generally defined as described therein. In addition, the general principles of Organic Chemistry, as well as specific functional moieties and reactivity, are described in "Organic Chemistry", Thomas Sorrell, University Science Books, Sausaltito: 1999, the entire content of which is incorporated herein by reference.

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

According to the present invention, isomeric mixtures containing any of a variety of isomer ratios may be used. For example, when only two isomers are mixed, 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. One of ordinary skill in the art will readily appreciate that similar ratios are expected for more complex mixtures of isomers.

The invention also includes isotopically-labeled compounds, which are identical to those disclosed herein, except 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 compounds of the invention include isotopes of hydrogen, carbon, nitrogen, oxygen, phosphorus, sulfur, fluorine and chlorine, such as²H、³H、¹³C、¹¹C、¹⁴C、¹⁵N、¹⁸O、¹⁷O、³¹P、³²P、³⁵S、¹⁸F and³⁶and (4) Cl. Comprises the aboveIsotopes of the compounds of the present invention or enantiomers, diastereomers, tautomers, or pharmaceutically acceptable salts or solvates thereof of other isotopes and/or other isotopes of other atoms 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, are useful in drug and/or substrate tissue distribution assays. Tritium-labelled i.e³H and carbon-14 i.e¹⁴The C isotope is particularly preferred because of its ease of preparation and detection. Also, with heavier isotopes such as deuterium²H substitution may provide therapeutic advantages resulting from greater metabolic stability, such as increased in vivo half-life or reduced dosage requirements, and may therefore be preferred in some circumstances. Isotopically labeled compounds can generally be prepared by carrying out the procedures disclosed in the schemes below and/or in the examples 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 afford the pure desired enantiomer. Alternatively, where the molecule contains a basic functional group such as an amino group or an acidic functional group such as a carboxyl group, diastereomeric salts are formed with an appropriate optically active acid or base, and the diastereomers thus formed are resolved by fractional crystallization or chromatographic means well known in the art, and the pure enantiomers are subsequently recovered.

It is to be understood that the compounds 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 the substituents contained in the formulae herein, refers to the replacement of a hydrogen in a given structure with the indicated substituent. When more than one position in any given structure may be substituted with more than one substituent selected from a specified group, the substituents at each position may be the same or different. As used herein, the term "substituted" is intended 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 which satisfy the valences of the heteroatom. In addition, 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 in the treatment, for example, of infectious diseases or proliferative disorders. As used herein, the term "stable" preferably refers to a compound that possesses sufficient stability to allow preparation and sufficient time to maintain the integrity of the compound for detection and preferably for a sufficient period of time to be useful for the purposes detailed herein.

Compound (I)

The novel cyclosporin derivatives of the present invention are potent inhibitors 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:

R₈is n-butyl, (E) -but-2-enyl or

R₂Is ethyl, 1-hydroxyethyl, isopropyl or n-propyl;

w is O, S or NR₁；

R₁Is hydrogen;

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

(C₃-C₇) Cycloalkyl, optionally with (C)₁-C₆) Alkyl substitution;

R₃the method comprises the following steps:

H；

(C₂-C₆) Alkynyl, optionally substituted by a group which may be the same or different selected from halogen, hydroxy, amino, monoalkylamino and dialkylOne or more groups of amino substituted;

R₇is that

R₅The method comprises the following steps:

H；

(C₂-C₆) AlkynesOptionally substituted with one or more groups which may be the same or different selected from halogen, hydroxy, amino, monoalkylamino and dialkylamino;

each occurrence of R_AAnd R_BIndependently are:

hydrogen;

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

(C₃-C₇) Cycloalkyl, optionally with (C)₁-C₆) Alkyl substitution;

each occurrence of R_CIndependently is hydrogen or (C)₁-C₆) An alkyl group;

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

m is an integer of 1, 2, 3, 4 or 5.

In certain embodiments, R₈Is n-butyl. In certain other embodiments, R₈Is (E) -but-2-enyl. In certain other embodiments, R₈Is thatIn certain embodiments, wherein R is₂Is ethyl.

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

or a pharmaceutically acceptable salt thereof, wherein:represents a single or double bond;

each W is independently O, S or NR₁；

Each R₁Independently is hydrogen;

(C₁-C₆) Alkyl, optionally substituted by one or more, which may be the same or differentA group R_DSubstitution;

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

(C₃-C₇) Cycloalkyl, optionally with (C)₁-C₆) Alkyl substitution;

each R₃Independently are:

H；

each R₅Independently are:

H；

each occurrence of R_AAnd R_BIndependently are:

hydrogen;

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

(C₃-C₇) Cycloalkyl, optionally with (C)₁-C₆) Alkyl substitution;

a phenyl group,optionally with a halogen atom which may be the same or different, selected from the group consisting of-O (C)₁-C₆) Alkyl, -C (═ O) O (C)₁-C₆) One to five groups of alkyl, amino, alkylamino and dialkylamino;

Each occurrence of R_CIndependently is hydrogen or (C)₁-C₆) An alkyl group;

each occurrence of R_FAnd R_F’Independently of each other, 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 three to seven ring atoms, which ring may optionally contain another heteroatom selected from the group consisting of nitrogen, oxygen and sulfur, and may optionally be substituted by one to four groups selected from the group consisting of alkyl, phenyl and benzyl, which may be the same or different;

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

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

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

In certain embodiments, W is O. In certain other embodiments, W is S. In yet other embodiments, W is NH. In still other embodiments, W is NR₁. In still other embodiments, W is N- (C)₁-C₄) An alkyl group.

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

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

In 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 of 2, 3, 4, 5 or 6; and wherein R is at each occurrence_AAnd R_BIndependently is hydrogen; (C)₁-C₄) Alkyl, optionally substituted by one or more radicals R which may be the same or different_DSubstituted, wherein R is at each occurrence_DIndependently 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 three to seven ring atoms, which ring may optionally contain another heteroatom selected from the group consisting of nitrogen, oxygen and sulfur and may optionally be substituted by one to four groups selected from (C) which may be the same or different (C)₁-C₄) Alkyl, phenyl and benzyl.

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

In certain embodiments, n is 2. In certain other embodiments, n is 3. In still other embodiments, n is 4, 5, or 6.

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-iso-butylaminoethyl, ethyl-iso-Butylaminoethyl, methyl-tert-butylaminoethyl or ethyl-tert-butylaminoethyl.

In certain embodiments, R₃Is thatWherein 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 still 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, W is NR₁And R is₁And R₃Together 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 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 one to three substituents of halogen; 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 of 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 still other embodiments, R₅Is CH₂-S-(C₁-C₆) Alkyl radicals, e.g. CH₂-S-CH₃. In still other embodiments, R₅Is CH₂-O-(C₁-C₆) Alkyl radicals, e.g. CH₂-O-CH₂-CH₃. In still other embodiments, R₅Is (C)₂-C₆) Alkenyl, e.g. CH₂-CH＝CH₂. In still other embodiments, R₅Is benzyl. In still other embodiments, R₅Is (C)₂-C₆) And (5) OH. In still other embodiments, R₅Is (C)₁-C₆) Monoalkylamines, e.g. CH₂-NH-Me. In still other embodiments, R₅Is (C)₁-C₆) Dialkylamines, e.g. CH₂-CH₂-N(Et)₂. In still other embodiments, R₅Is (C)₁-C₆) Cyclic amines, e.g. CH₂-CH₂-morpholine.

In certain embodiments, each occurrence of R_AAnd R_BIndependently 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 certain embodiments, whereinRepresents a single bond. In certain other embodiments, whereinRepresents a double bond.

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

or a pharmaceutically acceptable salt thereof, wherein:

represents a single or double bond;

each W is independently O, S or NR₁；

Each R₁Independently is hydrogen;

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

(C₃-C₇) Cycloalkyl, optionally with (C)₁-C₆) Alkyl substitution;

each R₅Independently are:

H；

(C₂-C₆) Alkenyl, optionally substituted by one or more substituents which may be the same or different, 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);

each occurrence of R_AAnd R_BIndependently are:

hydrogen;

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

(C₃-C₇) Cycloalkyl, optionally with (C)₁-C₆) Alkyl substitution;

or R_AAnd R_BTogether with the nitrogen atom to which they are attached, form a saturated or unsaturated heterocyclic ring containing three to seven ring atoms, which ring may optionally contain another heteroatom selected from the group consisting of nitrogen, oxygen and sulfur and may optionally be selected from the group consisting of identical or differentOne to four groups of the group consisting of alkyl, phenyl and benzyl;

each occurrence of R_CIndependently is hydrogen or (C)₁-C₆) An alkyl group;

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

Each of m, n, and q is independently an integer of 1, 2, 3, 4, or 5.

In another aspect, the invention provides compounds of formulae (IIb) - (Vb):

or a pharmaceutically acceptable salt thereof, wherein:

represents a single or double bond;

each R₁Independently is hydrogen;

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

(C₃-C₇) Cycloalkyl, optionally with (C)₁-C₆) Alkyl substitution;

or R₁And R₃Together with the nitrogen atom to which they are attached form a saturated or unsaturated heterocyclic ring containing three to seven ring atoms, which ring optionally contains another heteroatom selected from the group consisting of nitrogen, oxygen and sulfur and may optionally be selected from (C) which may be the same or different₁-C₆) One to four groups of the group consisting of alkyl, phenyl and benzyl;

each R₃Independently are:

H；

(C₂-C₆) Alkenyl, optionally substituted by one or more substituents which may be the same or different, 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_AOne or more ofSubstituted by one group;

each R₅Independently are:

H；

each occurrence of R_AAnd R_BIndependently are:

hydrogen;

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

(C₃-C₇) Cycloalkyl, optionally with (C)₁-C₆) Alkyl substitution;

or R_AAnd R_BTogether with the nitrogen atom to which they are attached, form a saturated or unsaturated heterocyclic ring containing three to seven ring atoms, which ring may optionally contain another heteroatom selected from the group consisting of nitrogen, oxygen and sulfur and may optionally be substituted by one to four groups which may be the same or different selected from the group consisting of alkyl, phenyl and benzyl;

each occurrence of R_CIndependently is hydrogen or (C)₁-C₆) An alkyl group;

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

Each of m, n, and q is independently an integer of 1, 2, 3, 4, or 5.

or a pharmaceutically acceptable salt thereof, wherein:

represents a single or double bond;

each W is independently O, S or NR₁；

Each R₁Independently is hydrogen;

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

(C₃-C₇) Cycloalkyl, optionally with (C)₁-C₆) Alkyl substitution;

or a heterocyclic ring, which may be saturated or unsaturated, containing five or six ring atoms and one to three heteroatoms, which may be the same or different, selected from nitrogen, sulfur and oxygen; each R₃Independently is

Each R₅Independently are:

H；

each occurrence of R₆Independently 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 a group which may be the same or differentSelected 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 occurrence of R_AAnd R_BIndependently are:

hydrogen;

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

(C₃-C₇) Cycloalkyl, optionally with (C)₁-C₆) Alkyl substitution;

each occurrence of R_CIndependently is hydrogen or (C)₁-C₆) An alkyl group;

each time goes outAt the occurrence of R_DIndependently 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; and is

Each of m, n, and q is independently an integer of 1, 2, 3, 4, or 5.

In certain embodiments, q is 1. In certain other embodiments, q is 2.

In certain embodiments, W is S. In certain other embodiments, W is O. In yet other embodiments, W is NH. In still other embodiments, W is N- (C)₁-C₄) An alkyl group.

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, R₁And R₃Together 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 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, each occurrence of R_AAnd R_BIndependently 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, each occurrence of R_AAnd R_BIndependently is H or (C)₁-C₆) An alkyl group. In still 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 one aspect, the present invention provides a compound selected from the group consisting of:

or a pharmaceutically acceptable salt thereof.

In a further aspect, the invention provides a compound as described in the examples.

In certain embodiments, the compound is selected from:

[ (R) - (2- (N, N-diethylamino) ethylthio) methyl-Sar ] -3- [ (gamma-hydroxy) -N-MeLeu ] -4-Cyclosporin,

[ (R) - (2- (N-ethyl-N-isopropylamino) ethylthio) methyl-Sar ] -3- [ (gamma-hydroxy) -N-MeLeu ] -4-cyclosporin,

[ (R) - (2- (N-isobutylamino) ethylthio) methyl-Sar ] -3- [ (gamma-hydroxy) -N-MeLeu ] -4-cyclosporin,

[ (R) - (2- (N-ethyl-N-isobutylamino) ethylthio) methyl-Sar ] -3- [ (gamma-hydroxy) -N-MeLeu ] -4-cyclosporin,

[ (R) - (2- (N-isobutyl-N-methylamino) ethylthio) methyl-Sar ] -3- [ (gamma-hydroxy) -N-MeLeu ] -4-cyclosporin,

[ (R) - (2- (N-neopentylamino) ethylthio) methyl-Sar ] -3- [ (gamma-hydroxy) -N-MeLeu ] -4-Cyclosporin,

[ (R) - (2- (N-methyl-N-neopentylamino) ethylthio) methyl-Sar ] -3- [ (gamma-hydroxy) -N-MeLeu ] -4-cyclosporin,

[ (R) - (2- (N-ethyl-N-neopentylamino) ethylthio) methyl-Sar ] -3- [ (gamma-hydroxy) -N-MeLeu ] -4-cyclosporin,

[ (R) - (2- (N-piperidinyl) ethylthio) methyl-Sar ] -3- [ (gamma-hydroxy) -N-MeLeu ] -4-cyclosporin,

[ (R) - (2- (N-morpholinyl) ethylthio) methyl-Sar ] -3- [ (gamma-hydroxy) -N-MeLeu ] -4-Cyclosporin,

[ (R) - (2- (N-thiomorpholinyl) ethylthio) methyl-Sar ] -3- [ (gamma-hydroxy) -N-MeLeu ] -4-Cyclosporin,

[ (R) - (2- (4-methyl-N-piperazinyl) ethylthio) methyl-Sar ] -3- [ (γ -hydroxy) -N-MeLeu ] -4-cyclosporin,

[ (R) - (3- (N, N-diethylamino) propylthio) methyl-Sar ] -3- [ (gamma-hydroxy) -N-MeLeu ] -4-cyclosporin,

[ (R) - (3- (N-ethyl-N-isopropylamino) propylthio) methyl-Sar ] -3- [ (gamma-hydroxy) -N-MeLeu ] -4-cyclosporin,

[ (R) - (3- (N-neopentylamino) propylthio) methyl-Sar ] -3- [ (gamma-hydroxy) -N-MeLeu ] -4-cyclosporin,

[ (R) - (3- (N-pyrrolidinyl) propylthio) methyl-Sar ] -3- [ (gamma-hydroxy) -N-MeLeu ] -4-cyclosporin,

[ (R) - (3- (N-piperidinyl) propylthio) methyl-Sar ] -3- [ (gamma-hydroxy) -N-MeLeu ] -4-cyclosporin,

[ (R) - (3- (N-morpholinyl) propylthio) methyl-Sar ] -3- [ (gamma-hydroxy) -N-MeLeu ] -4-cyclosporin,

[ (R) - (3- (N-thiomorpholinyl) propylthio) methyl-Sar ] -3- [ (gamma-hydroxy) -N-MeLeu ] -4-Cyclosporin,

[ (R) - (3- (4-methyl-N-piperazinyl) propylthio) methyl-Sar ] -3- [ (gamma-hydroxy) -N-MeLeu ] -4-cyclosporin,

[ (R) - (4- (N, N-diethylamino) butylthio) methyl-Sar ] -3- [ (gamma-hydroxy) -N-MeLeu ] -4-cyclosporin,

[ (R) - (4- (N-ethyl-N-isopropylamino) butylthio) methyl-Sar ] -3- [ (gamma-hydroxy) -N-MeLeu ] -4-cyclosporin,

[ (R) - (4- (N-isobutylamino) butylthio) methyl-Sar ] -3- [ (gamma-hydroxy) -N-MeLeu ] -4-cyclosporin,

[ (R) - (4- (N-isobutyl-N-methylamino) butylthio) methyl-Sar ] -3- [ (γ -hydroxy) -N-MeLeu ] -4-cyclosporin,

[ (R) - (4- (N-ethyl-N-isobutylamino) butylthio) methyl-Sar ] -3- [ (gamma-hydroxy) -N-MeLeu ] -4-cyclosporin,

[ (R) - (4- (N-neopentylamino) butylthio) methyl-Sar ] -3- [ (gamma-hydroxy) -N-MeLeu ] -4-cyclosporin,

[ (R) - (4- (N-methyl-N-neopentylamino) butylthio) methyl-Sar ] -3- [ (gamma-hydroxy) -N-MeLeu ] -4-cyclosporin,

[ (R) - (4- (N-piperidinyl) butylthio) methyl-Sar ] -3- [ (gamma-hydroxy) -N-MeLeu ] -4-cyclosporin,

[ (R) - (4- (N-morpholinyl) butylthio) methyl-Sar ] -3- [ (gamma-hydroxy) -N-MeLeu ] -4-cyclosporin,

[ (R) - (4- (N-thiomorpholinyl) butylthio) methyl-Sar ] -3- [ (gamma-hydroxy) -N-MeLeu ] -4-cyclosporin,

[ (R) - (4- (4-methyl-N-piperazinyl) butylthio) methyl-Sar ] -3- [ (gamma-hydroxy) -N-MeLeu ] -4-cyclosporin,

[ (R) - (3- (N-morpholinyl) propoxy) methyl-Sar ] -3- [ (gamma-hydroxy) -N-MeLeu ] -4-cyclosporin,

[ (R) - (2- (N, N-diethylamino) ethylthio) methyl-Sar ] -3- [ (gamma-methoxy) -N-MeLeu ] -4-cyclosporin,

[ (R) - (2- (N-pyrrolidinyl) ethylthio) methyl-Sar ] -3- [ (gamma-methoxy) -N-MeLeu ] -4-cyclosporin,

[ (R) - (3- (N, N-dimethylamino) propylthio) methyl-Sar ] -3- [ (gamma-methoxy) -N-MeLeu ] -4-cyclosporin,

[ (R) - (3- (N, N-diethylamino) propylthio) methyl-Sar ] -3- [ (gamma-methoxy) -N-MeLeu ] -4-cyclosporin,

[ (R) - (3- (N-piperidinyl) propylthio) methyl-Sar ] -3- [ (gamma-methoxy) -N-MeLeu ] -4-cyclosporin,

[ (R) - (4- (N-piperidinyl) butylthio) methyl-Sar ] -3- [ (gamma-methoxy) -N-MeLeu ] -4-cyclosporin,

[ (R) - (2- (N, N-diethylamino) ethoxy) methyl-Sar ] -3- [ (gamma-methoxy) -N-MeLeu ] -4-cyclosporin, and

[ (R) - (3- (N, N-diethylamino) propoxy) methyl-Sar ] -3- [ (γ -methoxy) -N-MeLeu ] -4-cyclosporin.

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

In a further aspect, the present invention provides a method of treating or preventing a viral infection in a mammalian species in need thereof, said method comprising administering to said 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 still other embodiments, the viral infection is an HCV infection. In still other embodiments, 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, said method comprising administering to said mammalian species a therapeutically effective amount of at least one compound described herein.

Preparation method

In certain embodiments, the compounds of formula (I) and (II) may be prepared by: treatment of cyclosporin A or an analogue thereof with a base (e.g., LDA) to form sarcosinate enolate at the 3-position followed by introduction of CO₂Gas to produce carboxylic acid-3-cyclosporin. After formation of its corresponding methyl ester and reduction of the methyl ester side chain to the alcohol, its mesylate or chloride may be produced by treatment with MsCl in dichloromethane solution. Both the mesylate and chloride salts can be converted to methylene groups on sarcosine by treatment with a base (e.g., NaH). When a 1, 4-addition reaction on a methylene group is carried out using a sulfur nucleophile, a methylene thioether chain with an R-conformation can be formed on the sarcosine in position 3 as a novel antiviral cyclosporin derivative. For example:

scheme 1

In certain embodiments, the methylene-3-cyclosporine obtained above may be converted to its methylene amine side chain to form a novel antiviral cyclosporine derivative. For example:

scheme 2

Thus, the following derivatives may be prepared according to the procedures described herein.

Scheme 3

In certain embodiments, the alcohol obtained above may be converted to its methylene oxy ether side chain to form a novel antiviral cyclosporin derivative. For example:

scheme 4

In schemes 1-4 above, the symbols have the same meaning as defined in the claims and throughout the specification, unless otherwise indicated.

In certain other embodiments, the compounds of formula (IIa) or (IIIa) may be obtained according to the procedures described herein.

Pharmaceutical composition

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

As used herein, the phrase "pharmaceutically acceptable carrier" refers to a pharmaceutically acceptable material, composition or vehicle, such as a liquid or solid filler, diluent, excipient, solvent or encapsulating material, involved in carrying or transporting the subject agent 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 materials that can 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; powdered tragacanth gum; 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; a phosphate buffer solution; and other non-toxic compatible materials employed in pharmaceutical formulations.

As set forth above, certain embodiments of the agents of the present invention may be provided in the form of pharmaceutically acceptable salts. In this respect, 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 may be prepared by separately reacting the purified compounds of the invention in free base form with a suitable organic or inorganic acid and isolating the salt thus formed. Representative salts include hydrobromide, hydrochloride, sulfate, bisulfate, phosphate, nitrate, acetate, valerate, oleate, palmitate, stearate, laurate, benzoate, lactate, phosphate, tosylate, citrate, maleate, fumarate, succinate, tartrate, naphylate, mesylate, glucoheptonate, lactobionate, and laurylate and the like. (see, e.g., Berge et al, (1977) "Pharmaceutical Salts", J.pharm.Sci.66: 1-19).

Pharmaceutically acceptable salts of the subject compounds include the conventional non-toxic salts or the quaternary ammonium salts of the compounds, for example, salts derived 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, butanoic 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, ethane disulfonic acid, oxalic acid, isethionic acid and the like.

In other cases, the compounds of the invention may contain one or more acidic functional groups and are therefore 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 likewise be prepared in situ during the final isolation and purification of the compounds, or can be prepared by reacting the purified compound in free acid form with a suitable base, such as the hydroxide, carbonate or bicarbonate of a pharmaceutically acceptable metal cation, alone or with ammonia or with a pharmaceutically acceptable organic primary, secondary or tertiary amine. Representative alkali or alkaline earth metal salts include lithium, sodium, potassium, calcium, magnesium, and aluminum and like salts. Representative organic amines useful for forming base addition salts include ethylamine, diethylamine, ethylenediamine, ethanolamine, diethanolamine, piperazine and similar organic amines. (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 may also be present in the composition.

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 can 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 a carrier material to produce a single dosage form will generally be that amount of compound which produces a therapeutic effect. Generally, the amount will range from about 1% to about 99% active ingredient, preferably from about 5% to about 70%, most preferably from about 10% to about 30% in 100%.

Methods of preparing these formulations or compositions include the step of bringing into association a compound of the invention with a carrier and optionally one or more accessory ingredients. In general, the formulations are prepared by: the compounds of the present invention are uniformly and intimately associated with a liquid carrier or a finely divided solid carrier or both, and the product is then shaped, if necessary.

Formulations of the invention suitable for oral administration may be in the form of: capsules, cachets, pills, tablets, lozenges (using a flavored base, usually 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 as a mouth wash 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 (capsules, tablets, pills, dragees, powders, granules and the like) for oral administration, the active ingredient is admixed 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; adsorbents such as kaolin and bentonite; 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 such excipients as lactose or milk sugar (milk sugars) as well as high molecular weight polyethylene glycols and similar excipients.

The tablets may be compressed or molded, optionally together with one or more accessory ingredients. Compressed tablets may employ binders (e.g., gelatin or hydroxybutyl methylcellulose), lubricants, inert diluents, preservatives, disintegrating agents (e.g., sodium starch glycolate or croscarmellose sodium), surface active agents or dispersing agents. Molded tablets may be prepared 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 formulating art. They may also be formulated to provide sustained or controlled release of the active ingredient therein using, for example, hydroxybutyl methylcellulose of different buttorions to provide the desired release profile, other polymer matrices, liposomes and/or microspheres. They can be sterilized by: for example, filtered bacteria retaining filters or by incorporating sterilizing agents in the form of sterile solid compositions that can be dissolved in sterile water or some other sterile injection medium just prior to use. These compositions may optionally also contain opacifying agents and may be of a composition that they release the active ingredient(s) only, or preferably, in a particular 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 microencapsulated form, if appropriate, using one or more of the excipients mentioned above.

Liquid dosage forms for oral administration of the compounds of the invention include pharmaceutically acceptable emulsions, microemulsions, solutions, suspensions, syrups and elixirs. 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. Additionally, cyclodextrins, such as hydroxybutyl- β -cyclodextrin, can be used to solubilize compounds.

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 hydroxide oxide (aluminum hydroxide), bentonite, agar-agar, and tragacanth, and mixtures thereof.

Formulations of the pharmaceutical compositions of the present invention for rectal or vaginal administration may be provided 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 will therefore 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 appropriate.

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 butellants that may be required.

In addition to the active compounds of the invention, ointments, pastes, creams, lotions and gels may contain excipients such as animal and vegetable fats, oils, waxes, paraffins, starch, tragacanth, cellulose derivatives, polyethylene glycols, silicones, bentonites, silicic acid, talc and zinc oxide, or mixtures thereof.

In addition to the compounds of the invention, powders and sprays may contain excipients such as lactose, talc, silicic acid, aluminum hydroxide, calcium silicates and polyamide powder or mixtures of these substances. Sprays can additionally contain customary butellants, such as chlorofluorohydrocarbons and volatile unsubstituted hydrocarbons, such as butane and butane.

Transdermal patches have the added 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 flow of the agents of the invention across the skin. The rate of such flow can be controlled by providing a rate controlling membrane or dispersing the compound in a polymer matrix or gel.

Ophthalmic formulations, eye ointments, powders, solutions, and the like are also contemplated as being 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 just 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, in order to prolong the effect of the drug, it is desirable to slow the absorption of the drug from subcutaneous or intramuscular injection. This can be achieved by using liquid suspensions of crystalline or amorphous materials that have poor water solubility. 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 oil vehicle. One strategy for depot injection involves the use of polyethylene oxide-polybutylene oxide copolymers where the vehicle is liquid at room temperature and solidifies at body temperature.

Injectable depot forms are prepared by forming a microencapsule matrix of the subject compound in a biodegradable polymer such as polylactic-polyglycolide. Depending on the ratio of drug to polymer and the nature of the particular polymer employed, the rate of release of the drug can be controlled. Other examples of biodegradable polymers include poly (orthoesters) and poly (anhydrides). Depot injectable formulations are also 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 as such 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, i.e., the compounds and pharmaceutical compositions may be administered concurrently with, before, or after one or more other desired therapeutic agents or medical procedures. The particular combination of therapies (therapeutic agents or procedures) employed in a combination regimen will take into account the compatibility of the desired therapeutic agent and/or procedure and the desired therapeutic effect to be achieved. It is also understood that the therapies employed can achieve the desired effect on the same condition (e.g., the compounds of the invention can be administered concurrently with another anti-HCV agent), or they can achieve different effects (e.g., control of any adverse effects).

The compounds of the 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 domestic 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 such containers may be accompanied by a notice in the form of a regulatory agency regulating the manufacture, use or sale of the drug or biological product, which notice reflects approval by the agency of manufacture, use or sale for human administration.

Equivalents of

The following representative examples are intended to help illustrate the invention, and are not intended to, nor should they be construed as, limiting the scope of the invention. Indeed, various modifications of the invention and many further embodiments thereof will be apparent to those skilled in the art in light of the entire contents of this document, including the following examples and references to scientific and patent documents cited therein, in addition to those shown and described herein. It is to be further understood that the contents of those cited references are incorporated herein by reference to help clarify the background. The following examples contain important additional information, exemplification and guidance which can be adapted to the implementation of the various embodiments of the invention and their equivalents.

Examples

Example 1

[ alpha-methoxycarbonyl-Sar ] -3-cyclosporin

Reacting [ alpha-carboxy-sar]-3-Cyclosporin (5.00g, 4.01mmol) was dissolved in 30ml of N, N-dimethylformamide. 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. Then 60ml of ethyl acetate and 60ml of water were 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 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)⁺；TLC R_f: 0.55 (dichloromethane/methanol ═ 9/1)]。

Example 2

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

Reacting [ alpha-methoxycarbonyl-Sar ]]-3-Cyclosporin (2.00g, 1.59mmol) was dissolved in tetrahydrofuran (30 ml). Cesium chloride (1.33g) and sodium borohydride (0.60g, 15.89mmol) were added portionwise. To the mixture was added dropwise 30ml of methanol over 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 yield 1.99g of crude product which was purified by column on silica gel with dichloromethane/methanol (100: 0 to 95: 5) to yield 1.50g of pure product (yield: 76%) [ formula: c₆₃H₁₁₃N₁₁O₁₃(ii) a Accurate quality: 1231.85, respectively; MS (m/z): 1232.7(M +1)⁺、1254.7(M+Na)⁺]。

Example 3

[ alpha-Methylmethanesulfonate-Sar ] -3-Cyclosporin

At 0 deg.C to [ alpha-hydroxymethyl-Sar ]]-3-Cyclosporium spA solution of the antibiotic (30mg, 0.024mmol) in dichloromethane (2ml) was added triethylamine (52.8. mu.l, 0.38mmol) and methanesulfonyl chloride (23mg, 0.20 mmol). The mixture was stirred at room temperature for two hours. The reaction mixture was then washed with brine, dried over magnesium sulfate and evaporated under reduced pressure to yield 33mg of crude product, which was used in the next reaction without further purification [ formula: c₆₄H₁₁₅N₁₁O₁₅S; accurate quality: 1309.83, respectively; MS (m/z): 1310.7(M +1)⁺]。

Example 4

[ alpha-chloromethyl-Sar ] -3-cyclosporin

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

Example 5

[ alpha-methylene-Sar ] -3-Cyclosporin

At 0 deg.C to [ alpha-methanesulfonate methyl-Sar]-3-Cyclosporin (33mg, 0.025mmol) or [ alpha-chloromethyl-Sar]A solution of-3-cyclosporin (30mg, 0.025mmol) in tetrahydrofuran (3ml) was added sodium hydride (15.3mg, 60% in oil, 0.38mmol, 10 equiv). The mixture was stirred at 0 ℃ for one hour, then warmed to room temperature for 30 minutes. After removal of the solvent, the residue was dissolved in 20ml of dichloromethane. 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 using dichloromethane/methanol (20/1) to yield 16mg of the 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)]。

Example 6

[ (R) - (2- (N, N-dimethylamino) ethylthio) methyl-Sar ] -3-cyclosporin (isomer B) and [ (S) - (2- (N, N-dimethylamino) ethylthio) methyl-Sar ] -3-cyclosporin (isomer A)

To [ alpha-methylene-Sar ]]A solution of-3-cyclosporin (0.60g, 0.50mmol) and 2- (dimethylamino) ethanethiol (0.63g, 6.00mmol) in methanol (20ml) was added triethylamine (0.82ml, 6.0 mmol). The reaction mixture was stirred at room temperature overnight. After removal of the solvent, the residue was subjected to chromatography using methylene/methanol as eluent to give 0.35g of (S) -2- (N, N-dimethylamino) ethylthiomethyl-Sar]-3-Cyclosporin (isomer A) and 0.20g of [ (R) -2- (N, N-dimethylamino)) Ethyl thiomethyl-Sar]-3-cyclosporin (isomer B) [ formula: c₆₇H₁₂₂N₁₂O₁₂S; accurate quality: 1218.9, respectively; MS (m/z): 1319.80(M +1)⁺；TLC R_f: 0.20 (ethyl acetate/methanol ═ 5/1); HPLC RT: 12.55min. (isomer A) and 13.22min. (isomer B) (C8 reverse phase column: 250mm, acetonitrile/water (0.05% trifluoroacetic acid), operating temperature: 64 ℃ C.; detector: 210nm)]。

Example 7

[ (R) - (2- (N, N-diethylamino) ethylthio) methyl-Sar ] -3-cyclosporin (isomer B) and [ (S) - (2- (N, N-diethylamino) ethylthio) methyl-Sar ] -3-cyclosporin (isomer A)

To [ alpha-methylene-Sar ]]A solution of-3-cyclosporin (0.31g, 0.25mmol) and 2-diethylaminoethanethiol (0.40g, 3.00mmol) in methanol (10ml) was added triethylamine (0.41ml, 3.00mmol, 12 equivalents). The reaction mixture was stirred at room temperature overnight. After removal of the solvent, the residue was subjected to chromatography using methylene/methanol as eluent to give 0.20g of [ (S) -2- (N, N-diethylamino) ethylthiomethyl-Sar]-3-Cyclosporin (isomer A) and 0.08g of [ (R) -2- (N, N-diethylamino) ethylthiomethyl-Sar]-3-cyclosporin (isomer B) [ formula: c₆₉H₁₂₆N₁₂O₁₂S; accurate quality: 1346.93, respectively; MS (m/z): 1347.80(M +1)⁺；TLC R_f: 0.23 (ethyl acetate/methanol ═ 5/1); HPLC RT: 13.37min. (isomer A) and 13.91min. (isomer B) (C8 reverse phase column: 250mm, acetonitrile/water (0.05% trifluoroacetic acid), operating temperature: 64 ℃ C.; detector: 210nm)]。

Example 8

[ (R) - (2- (N, N-diethylamino) ethoxy) methyl-Sar ] -3-cyclosporin

To [ (R) -alpha-hydroxymethyl-Sar]A solution of-3-cyclosporin (0.36g, 0.29mmol) in benzene (30ml) was added a solution of sodium hydroxide (1.20g, 30mmol) in water (2ml), 2-bromo-N, N-diethylethanamine hydrobromide (3.80g, 14.56mmol) and tetra-N-butylammonium bromide (0.20g, 0.62 mmol). The reaction mixture was stirred at 30 ℃ for 20 hours. After dilution with ice water, the mixture was separated. The aqueous layer was extracted with dichloromethane (30 ml). The combined organic layers were washed with brine, dried over magnesium sulfate and evaporated under reduced pressure. The residue was purified by chromatography (dichloromethane/methanol 96/4) to yield 210mg of product [ formula: c₆₉H₁₂₆N₁₂O₁₃(ii) a Accurate quality: 1330.96, respectively; MS (m/z): 1331.71(M +1)⁺；TLC R_f: 0.38 (dichloromethane/methanol 95/5); HPLC RT: 14.12min (C8 reverse phase column: 250 mm; acetonitrile/0.077% ammonium acetate in water; operating temperature: 64 ℃ C.; detector: 210nm)]。

Example 9

[ (R) -alpha- (tert-butoxycarbonylmethoxy) methyl-Sar ] -3-cyclosporin

To [ (R) -alpha-hydroxymethyl-Sar]A solution of-3-Cyclosporin (0.50g, 0.41mmol) in benzene (30ml) was added a solution of sodium hydroxide (1.00g, 25.00mmol) in water (1ml), tert-butyl bromoacetate (3.20g, 16.41mmol) and tetra-n-butylammonium bromide (0.40g, 1.24 mmol)mmol). The mixture was stirred at room temperature for 10 hours. After dilution with ice water, the mixture was separated. The aqueous layer was extracted with dichloromethane (30 ml). The combined organic layers were washed with brine, dried over magnesium sulfate and evaporated under reduced pressure. The residue was purified by chromatography (hexane/acetone 2/1) to yield 0.41g of product [ formula: c₆₉H₁₂₃N₁₁O₁₅(ii) a Accurate quality: 1345.92, respectively; MS (m/z): 1346.61(M +1)⁺；TLC R_f: 0.60 (dichloromethane/methanol 95/5); HPLC RT: 18.29min (C8 reverse phase column: 250 mm; acetonitrile/0.077% ammonium acetate in water; operating temperature: 64 ℃ C.; detector: 210nm)]。

Example 10

[ (R) -alpha- (ethoxycarbonylmethoxy) methyl-Sar ] -3-cyclosporin

To [ (R) -alpha-hydroxymethyl-Sar]A solution of-3-cyclosporin (0.35g, 0.28mmol) in benzene (15ml) was added a solution of sodium hydroxide (0.60g, 15.00mmol) in water (1ml), ethyl bromoacetate (1.60g, 9.58mmol) and tetra-n-butylammonium bromide (0.20g, 0.62 mmol). The mixture was stirred at room temperature for 10 hours. After dilution with ice water, the mixture was separated. The aqueous layer was extracted with dichloromethane (15 ml). The combined organic layers were washed with brine, dried over magnesium sulfate and evaporated under reduced pressure. The residue was purified by chromatography (hexane/acetone 2/1) to yield 0.31g of product [ formula: c₆₇H₁₁₉N₁₁O₁₅(ii) a Accurate quality: 1317.89, respectively; MS (m/z): 1318.46(M +1)⁺；TLC R_f: 0.55 (dichloromethane/methanol 95/5); HPLCRT: 17.40min (C8 reverse phase column: 250 mm; acetonitrile/0.077% ammonium acetate in water; operating temperature: 64 ℃ C.; detector: 210nm)]。

Example 11

[ (R) -alpha- (carboxymethoxy) methyl-Sar ] -3-cyclosporin

To [ (R) -alpha- ((tert-butoxycarbonyl) methoxy) methyl-Sar](iii) -3-Cyclosporin (0.18g, 0.13mmol) in 5ml dichloromethane trifluoroacetic acid (1ml) and Et₃SiH (10 drops). The mixture was stirred at room temperature for 3 hours and concentrated under reduced pressure. Dichloromethane (10ml) and water (10ml) were then added and the mixture was separated. The dichloromethane layer was dried over magnesium sulfate and evaporated under reduced pressure. The residue was purified by C-18 chromatography (acetonitrile/water) to yield 75mg of product [ formula: c₆₅H₁₁₅N₁₁O₁₅(ii) a Accurate quality: 1289.86, respectively; MS (m/z): 1290.56(M +1)⁺(ii) a HPLC RT: 11.03min (C8 reverse phase column: 250 mm; acetonitrile/water (0.05% trifluoroacetic acid); operating temperature: 64 ℃ C.; detector: 210nm)]。

Example 12

[ (R) -alpha- (carboxymethoxy) methyl-Sar ] -3-cyclosporin-sodium salt

To [ (R) -alpha- (carboxymethoxy) methyl-Sar]A solution of-3-cyclosporin (30mg, 0.02mmol) in methanol (1ml) was added a solution of sodium hydroxide (1.00mg, 0.02mmol) in water (0.5 ml). The mixture was stirred at room temperature for 1 hour and dried under high vacuum to give 28mg of product [ formula: c₆₅H₁₁₄N₁₁NaO₁₅(ii) a Accurate quality: 1311.84；MS(m/z)：1290.56(M+1-Na)⁺(ii) a HPLC RT: 10.98min (C8 reverse phase column: 250 mm; acetonitrile/water (0.05% trifluoroacetic acid); operating temperature: 64 ℃ C.; detector: 210nm)]。

Example 13

[ (R) - (2- (N, N-dimethylamino) ethoxy) methyl-Sar ] -3-cyclosporin

To [ (R) -alpha-hydroxymethyl-Sar]A solution of-3-cyclosporin (1.03g, 0.84mmol) in benzene (50ml) was added a solution of sodium hydroxide (1.34g, 33.47mmol) in water (1.34ml), tetramethylammonium hydroxide pentahydrate (3.04g, 16.73mmol) and 2-dimethylaminoethyl chloride hydrochloride (2.41g, 16.73 mmol). The mixture was stirred at room temperature for 5 days. A saturated solution of sodium bicarbonate (100ml) was added and the mixture was separated. The aqueous layer was then extracted with ethyl acetate (50 ml. times.2). The combined organic layers were dried over magnesium sulfate and evaporated under reduced pressure. After purification on silica gel, 303mg of product [ formula: c₆₇H₁₂₂N₁₂O₁₃(ii) a Accurate quality: 1302.93, respectively; MS (m/z): 1303.70(M +1)⁺、1325.85(M+Na)⁺；TLC R_f: 0.36 (dichloromethane/methanol ═ 9/1); HPLC RT: 18.19min (C8 reverse phase column: 250 mm; acetonitrile/0.077% ammonium acetate in water; operating temperature: 64 ℃ C.; detector: 210nm)]。

Example 14

[ (R) - (2- (N-morpholinyl) ethoxy) methyl-Sar ] -3-cyclosporin

To [ (R) -alpha-hydroxymethyl-Sar]A solution of-3-cyclosporin (0.27g, 0.22mmol) in benzene (20ml) was added a solution of sodium hydroxide (0.70g, 17.55mmol) in water (0.70ml), tetramethylammonium hydroxide pentahydrate (0.80g, 4.39mmol) and 2- (4-morpholinyl) ethylchloride hydrochloride (0.82g, 4.39 mmol). The mixture was stirred at 30 to 40 ℃ for one week. A saturated solution (30ml) of sodium hydrogencarbonate was added, and the mixture was separated. The aqueous layer was extracted with ethyl acetate (25 ml. times.2). The combined organic layers were dried over magnesium sulfate and evaporated under reduced pressure. After purification on silica gel, 56mg of product are obtained [ formula: c₆₉H₁₂₄N₁₂O₁₄(ii) a Accurate quality: 1344.94, respectively; MS (m/z): LCMS: 1345.72(M +1)⁺、1367.83(M+Na)⁺；TLC R_f: 0.50 (dichloromethane/methanol ═ 9/1); HPLC RT: 16.64min (C8 reverse phase column: 250 mm; acetonitrile/0.077% ammonium acetate in water; operating temperature: 64 ℃ C.; detector: 210nm)]。

Example 15

[ (R) - (2- (N-pyrrolidinyl) ethoxy) methyl-Sar ] -3-cyclosporin

To [ (R) -alpha-hydroxymethyl-Sar]A solution of-3-cyclosporin (0.320g, 0.26mmol) in benzene (20ml) was added a solution of sodium hydroxide (0.83g, 20.80mmol) in water (0.85ml), tetramethylammonium hydroxide pentahydrate (0.95g, 5.20mmol) and 1- (2-chloroethyl) pyrrolidine hydrochloride (0.88g, 5.20 mmol). The mixture was stirred at room temperature over the weekend. A saturated solution of sodium bicarbonate (30ml) was added and the mixture was separated. The aqueous layer was extracted with ethyl acetate (25 ml. times.2). The combined organic layers were dried over magnesium sulfate and evaporated under reduced pressure. After purification on silica gel, 103mg of product [ formula：C₆₉H₁₂₄N₁₂O₁₃(ii) a Accurate quality: 1328.94, respectively; MS (m/z): 1329.75(M +1)⁺、1351.82(M+Na)⁺；TLC R_f: 0.37 (dichloromethane/methanol ═ 9/1); HPLC RT: 18.94min (C8 reverse phase column: 250 mm; acetonitrile/0.077% ammonium acetate in water; operating temperature: 64 ℃ C.; detector: 210nm)]。

Example 16

[ (R) - (2- (N-piperidinyl) ethoxy) methyl-Sar ] -3-cyclosporin

To [ (R) -alpha-hydroxymethyl-Sar]A solution of-3-cyclosporin (0.28g, 0.22mmol) in benzene (20ml) was added a solution of sodium hydroxide (0.36g, 9.07mmol) in water (0.36ml), tetramethylammonium hydroxide pentahydrate (0.82mg, 4.53mmol) and 1- (2-chloroethyl) piperidine (piperidine) hydrochloride (0.83g, 4.53 mmol). The mixture was stirred at 30 to 40 ℃ for 20 hours. A saturated solution of sodium hydrogencarbonate (30ml) was added and the mixture was separated. The aqueous layer was then extracted with ethyl acetate (25 ml. times.2). The combined organic layers were dried over magnesium sulfate and evaporated under reduced pressure. After purification on silica gel, 121mg of product are obtained [ formula: c₇₀H₁₂₆N₁₂O₁₃(ii) a Accurate quality: 1342.96, respectively; MS (m/z): 1343.76(M +1)⁺、1365.83(M+Na)⁺；TLC R_f: 0.44 (dichloromethane/methanol ═ 9/1); HPLC RT: 19.26min (C8 reverse phase column: 250 mm; acetonitrile/0.077% ammonium acetate in water; operating temperature: 64 ℃ C.; detector: 210nrm)]。

Example 17

[ (R) -alpha- (3, 3-dimethoxypropoxy) methyl-Sar ] -3-cyclosporin

To [ (R) -alpha-hydroxymethyl-Sar]A solution of-3-cyclosporin (0.50g, 0.41mmol) in benzene (30ml) was added a solution of sodium hydroxide (1.00g, 25.00mmol) in water (1ml), 3-bromopropionaldehyde dimethyl acetal (1.80g, 10.00mmol) and tetra-n-butylammonium bromide (0.20g, 0.62 mmol). After stirring at room temperature for 10 hours, the mixture was diluted with ice water and the mixture was separated. The aqueous layer was extracted with dichloromethane (20 ml). The combined organic layers were washed with brine, dried over magnesium sulfate and evaporated under reduced pressure to yield 0.48g of crude product, which was used in the next step [ formula: c₆₈H₁₂₃N₁₁O₁₅(ii) a Accurate quality: 1333.92, respectively; MS (m/z): 1334.50(M +1)⁺]。

Example 18

[ (R) -alpha- (2-formylethoxy) methyl-Sar ] -3-cyclosporin

To crude [ (R) - (3, 3-dimethoxypropoxy) methyl-Sar at 0 deg.C]A solution of-3-cyclosporin (0.48g, 0.36mmol) in dichloromethane (30ml) was added trifluoroacetic acid (5ml) and water (4 ml). The mixture was then allowed to warm to room temperature and stirred for 3 hours. After separation of the mixture, the dichloromethane layer was washed with saturated sodium bicarbonate solution (20ml), dried over magnesium sulfate and evaporated under reduced pressure. The residue was purified by chromatography (hexane/acetone 2/1) to yield 0.31g of product [ formula: c₆₆H₁₁₇N₁₁O₁₄(ii) a Accurate quality: 1287.88, respectively; MS (m/z): 1288.63(M +1)⁺]。

Example 19

[ (R) - (3- (N, N-dimethylamino) propoxy) methyl-Sar ] -3-cyclosporin

To [ (R) - (2-formylethoxy) methyl-Sar]A solution of-3-cyclosporin (0.13g, 0.10mmol) in chloroform (5ml) was added dimethylamine hydrochloride (0.10g, 1.22mmol) and acetic acid (5 drops). After the mixture was stirred at room temperature for 5 minutes, tetramethylammonium triacetoxyborohydride (65mg, 0.25mmol) was added in portions and stirring was continued for 1 hour. Dichloromethane (10ml) and saturated sodium bicarbonate solution (10ml) were then added and 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 96/4) to yield 89mg of product [ formula: c₆₈H₁₂₄N₁₂O₁₃(ii) a Accurate quality: 1316.94, respectively; MS (m/z): 1317.64(M +1)⁺；TLC R_f: 0.39 (dichloromethane/methanol 95/5); HPLC RT: 13.92min (C8 reverse phase column: 250 mm; acetonitrile/0.077% ammonium acetate in water; operating temperature: 64 ℃ C.; detector: 210nm)]。

Example 20

[ (R) - (3- (N, N-diethylamino) propoxy) methyl-Sar ] -3-cyclosporin

To [ (R) - (2-formylethoxy) methyl-Sar]A solution of-3-cyclosporin (100mg, 0.08mmol) in chloroform (4ml) was added diethylamine (100mg,1.37mmol) and acetic acid (4 drops). After the mixture was stirred at room temperature for 5 minutes, tetramethylammonium triacetoxyborohydride (50mg, 0.19mmol) was added in portions and stirring was continued for 1 hour. Dichloromethane (10ml) and saturated sodium bicarbonate solution (10ml) were then 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 96/4) to yield 56mg of product [ formula: c₇₀H₁₂₈N₁₂O₁₃(ii) a Accurate quality: 1344.97, respectively; MS (m/z): 1345.71(M +1)⁺；TLC R_f: 0.40 (dichloromethane/methanol 95/5); HPLC RT: 14.59min (C8 reverse phase column: 250 mm; acetonitrile/0.077% ammonium acetate in water; operating temperature: 64 ℃ C.; detector: 210nm)]。

Example 21

[ (R) - (3- (N-morpholinyl) propoxy) methyl-Sar ] -3-cyclosporin

To [ (R) - (2-formylethoxy) methyl-Sar]A solution of-3-cyclosporin (300mg, 0.23mmol) in dichloromethane (15ml) was added morpholine (101mg, 1.16mmol) and tetramethylammonium triacetoxyborohydride (306mg, 1.16 mmol). The reaction mixture was stirred at room temperature for two hours. Then a saturated solution of sodium bicarbonate (30ml) and dichloromethane (15ml) were added and the mixture was separated. The dichloromethane layer was dried over magnesium sulfate and evaporated under reduced pressure. After purification on silica gel, 124mg of product [ formula: c₇₀H₁₂₆N₁₂O₁₄(ii) a Accurate quality: 1358.95, respectively; MS (m/z): 1359.71(M +1)⁺、1381.79(M+Na)⁺；TLC R_f: 0.40 (dichloromethane/methanol ═ 9/1); HPLC RT: 14.2min (C8 reverse phase column: 250 mm; acetonitrile/0.077% ammonium acetate in water; operating temperature: 64 ℃ C.; detector: 210nm)]。

Example 22

[ (R) - (3- (N-pyrrolidinyl) propoxy) methyl-Sar ] -3-cyclosporin

To [ (R) -alpha-hydroxymethyl-Sar]A solution of-3-cyclosporin (315mg, 0.24mmol) in dichloromethane (15ml) was added pyrrolidine (87mg, 1.22mmol) and tetramethylammonium triacetoxyborohydride (322mg, 1.22 mmol). The reaction mixture was stirred at room temperature for 2 hours. Then a saturated solution of sodium bicarbonate (30ml) and dichloromethane (15ml) were added and the mixture was separated. The dichloromethane layer was dried over magnesium sulfate and evaporated under reduced pressure. After purification on silica gel, 22mg of product [ formula: c₇₀H₁₂₆N₁₂O₁₃(ii) a Accurate quality: 1342.96, respectively; MS (m/z): 1343.75(M +1)⁺、1365.82(M+Na)⁺；TLC R_f: 0.33 (dichloromethane/methanol ═ 9/1); HPLC RT: 14.3min (C8 reverse phase column: 250 mm; acetonitrile/0.077% ammonium acetate in water; operating temperature: 64 ℃ C.; detector: 210nm)]。

Example 23

[ (R) - (3- (N-piperidinyl) propoxy) methyl-Sar ] -3-cyclosporin

To [ (R) - (2-formylethoxy) methyl-Sar](350mg, 0.27mmol) of (E) -3-Cyclosporin in dichloromethane (20ml) piperidine (115mg, 1.34mmol) and tetramethyl triacetoxyborohydrideAmmonium (352mg, 1.34 mmol). The reaction mixture was stirred at room temperature overnight. Then a saturated solution of sodium bicarbonate (30ml) and dichloromethane (15ml) were added and the mixture was separated. The dichloromethane layer was dried over magnesium sulfate and evaporated under reduced pressure. After purification on silica gel, 35mg of product are obtained [ formula: c₇₁H₁₂₈N₁₂O₁₃(ii) a Accurate quality: 1356.97, respectively; MS (m/z): 1357.76(M +1)⁺、1379.83(M+Na)⁺；TLC R_f: 0.36 (dichloromethane/methanol ═ 9/1); HPLC RT: 14.4min (C8 reverse phase column: 250 mm; acetonitrile/0.077% ammonium acetate in water; operating temperature: 64 ℃ C.; detector: 210nm)]。

Example 24

[ alpha-carboxy-Sar ] -3- [ (gamma-hydroxy) -N-MeLeu ] -4-Cyclosporin

To a solution of LDA (2.0M in tetrahydrofuran, 23ml, 46mmol) in tetrahydrofuran (80ml) at-78 deg.C under nitrogen was added [ (gamma-hydroxy) -N-MeLeu ] in tetrahydrofuran (15ml) over 3min]-4-Cyclosporin (4.40g, 3.61 mmol). 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 allowed to warm slowly 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 around 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 yield 3.20g of crude product 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)⁺]。

[ (γ -hydroxy) -N-MeLeu ] -4-Cyclosporin was prepared by bioconversion from Sebekia benihana according to the method described by Kuhnt M. et al, 1996, Microbial Biotransformation Products of cyclosporine A, J.antibiotics, 49(8), 78.

Example 25

[ alpha-methoxycarbonyl-Sar ] -3- [ (gamma-hydroxy) -N-MeLeu ] -4-cyclosporin

To [ alpha-carboxy-Sar ]]-3- [ (γ -hydroxy) -N-MeLeu]A mixture of-4-cyclosporin (3.20g 2.53mmol) and potassium carbonate (1.30g, 9.40mmol) 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 crude 3.00g of the product [ formula: c₆₄H₁₁₃N₁₁O₁₅(ii) a Accurate quality: 1275.84, respectively; MS (m/z): 1276.75(M +1)⁺]。

Example 26

[ (R) -alpha-hydroxymethyl-Sar ] -3- [ (gamma-hydroxy) -N-MeLeu ] -4-cyclosporin

To [ alpha-methoxycarbonyl-Sar ]]-3- [ (γ -hydroxy) -N-MeLeu]A suspension of-4-cyclosporin (3.00g, 2.35mmol) and lithium chloride (1.50g, 35.30mmol) in methanol (100ml) was added portionwise to sodium borohydride (2.50g, 66.10 mmol). The mixture was stirred at room temperatureStirring 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 yield 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)]。

Example 27

[ alpha-methylene-Sar ] -3- [ (gamma-hydroxy) -N-MeLeu ] -4-Cyclosporin

[ alpha-methylene-Sar ]]-3- [ (γ -hydroxy) -N-MeLeu]-4-cyclosporin was prepared according to the method described in examples 4 and 5 [ 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)]。

Example 28

[ (R) - (2- (N, N-dimethylamino) ethylthio) methyl-Sar ] -3- [ (γ -hydroxy) -N-MeLeu ] -4-cyclosporin (isomer B) and [ (S) - (2- (N, N-dimethylamino) ethylthio) methyl-Sar ] -3- [ (γ -hydroxy) -N-MeLeu ] -4-cyclosporin (isomer A)

Reacting [ alpha-methylene-Sar]-3- [ (γ -hydroxy) -N-MeLeu]-4-Cyclosporin (0.62g, 0.50mmol) and 2- (dimethylamino) ethanethiol (0.49g, 6.00mmol) were dissolved in methanol (30ml) and triethylamine (0.82ml, 6.00mmol) was added. The mixture was stirred at room temperature overnight. After removal of the solvent, the residue was subjected to chromatography using dichloromethane/methanol as eluent, yielding 0.41g of [ (S) - (2- (N, N-dimethylamino) ethylthio) methyl-Sar [ ((S) - (2-N, N-dimethylamino) ethylthio) ] methyl-Sar []-3- [ (γ -hydroxy) -N-MeLeu]-4-Cyclosporin (isomer A) and 0.18g of [ (R) - (2- (N, N-bis)Methylamino) ethylthio) methyl-Sar]-3- [ (γ -hydroxy) -N-MeLeu]-4-cyclosporin (isomer B) [ formula: c₆₇H₁₂₂N₁₂O₁₃S; accurate quality: 1334.9, respectively; MS (m/z): 1335.7(M +1)⁺；TLC R_f: 0.05 (ethyl acetate/methanol ═ 5/1); HPLC RT: 10.88min. (isomer A) and 11.30min. (isomer B) (C8 reverse phase column: 250mm, acetonitrile/water (0.05% trifluoroacetic acid), operating temperature: 64 ℃ C.; detector: 210nm)]。

Example 29

[ (R) - (2- (N, N-diethylamino) ethylthio) methyl-Sar ] -3- [ (γ -hydroxy) -N-MeLeu ] -4-cyclosporin (isomer B) and [ (S) - (2- (N, N-diethylamino) ethylthio) methyl-Sar ] -3- [ (γ -hydroxy) -N-MeLeu ] -4-cyclosporin (isomer A)

Reacting [ alpha-methylene-Sar]-3- [ (γ -hydroxy) -N-MeLeu]-4-Cyclosporin (0.31g, 0.25mmol) and 2-diethylaminoethanethiol (0.40g, 3.00mmol) were dissolved in methanol (30ml), and triethylamine (0.41ml, 3.00mmol) was added. The mixture was stirred at room temperature overnight. After removal of the solvent, the residue was subjected to chromatography using dichloromethane/methanol as eluent, yielding 0.15g of [ (S) - (2- (N, N-diethylamino) ethylthio) methyl-Sar]-3- [ (γ -hydroxy-N-MeLeu)]-4-Cyclosporin (isomer A) and 0.10g of [ (R) - (2- (N, N-diethylamino) ethylthio) methyl-Sar]-3- [ (γ -hydroxy-N-MeLeu)]-4-cyclosporin (isomer B) [ formula: c₆₉H₁₂₆N₁₂O₁₃S; accurate quality: 1362.93, respectively; MS (m/z): 1363.75(M +1)⁺；TLC R_f: 0.1 (ethyl acetate/methanol ═ 5/1); HPLC RT: 11.64min. (isomer A) and 11.85min. (isomer B) (C8 reverse phase column: 250mm, acetonitrile/water (0.05% trifluoroacetic acid), operating temperature: 64 ℃ C.; detector: 210nm)]。

Example 30

[ (R) - (2- (N-morpholinyl) ethylthio) methyl-Sar ] -3- [ (γ -hydroxy) -N-MeLeu ] -4-Cyclosporin

To [ alpha-methylene-Sar ]]-3- [ (γ -hydroxy) -N-MeLeu]A solution of-4-cyclosporin (260mg, 0.21mmol) and 2-morpholinoethanethiol (300mg, 2.04mmol) in methanol (30ml) was added with lithium hydroxide (140mg, 5.83 mmol). The reaction mixture was stirred at room temperature overnight. Most of the solvent was evaporated under reduced pressure. Dichloromethane (30ml) and water (30ml) were added and the mixture was separated. The organic layer was washed with water and brine, dried over magnesium sulfate and evaporated under reduced pressure. The residue was purified by chromatography (dichloromethane/methanol 97/3) to yield 102mg of product [ formula: c₆₉H₁₂₄N₁₂O₁₄S; accurate quality: 1376.91, respectively; MS (m/z): 1399.85(M + Na)⁺(ii) a TLC Rf: 0.30 (dichloromethane/methanol ═ 9/1); HPLC RT: 11.03min (C8 reverse phase column: 250 mm; acetonitrile/0.077% ammonium acetate in water; operating temperature: 64 ℃ C.; detector: 210 nm);¹h NMR Spectrum (600MHz, CDCl)₃δ in ppm): 0.68(d, J ═ 6.6Hz, 3H), 0.79(d, J ═ 6.6Hz, 3H), 0.82(m, 6H), 0.85(d, J ═ 6.6Hz, 3H), 0.88(d, J ═ 7.2Hz, 3H), 0.90(d, J ═ 6.6Hz, 3H), 0.93(d, J ═ 6.6Hz, 3H), 0.97-1.00(m, 9H), 1.08(d, J ═ 6.6Hz, 3H), 1.21-1.25(m, 11H), 1.31(d, J ═ 7.2Hz, 3H), 1.39-1.47(m, 2H), 1.54-1.61(m, 8H), 1.66-1.70(m, 2H), 1.01(m, 2H), 2H (m, 3H), 3.7, 3H), 3.39-1.47 (m, 2H), 1.54-1.8H, 8H, 1.70(m, 3H), 3H, 3, 3H) 3.48(s, 3H), 3.52(br, 1H), 3.67(m, 6H), 4.51(m, 1H), 4.59(t, J ═ 8.4Hz, 1H), 4.81(m, 1H), 4.94-5.00(m, 2H), 5.04(t,J＝6.6Hz，1H)，5.08(d，J＝10.8Hz，1H)，5.27-5.31(m，1H)，5.33-5.37(m，1H)，5.48(m，2H)，5.67(m，1H)，7.14(d，J＝7.8Hz，1H)，7.49(d，J＝7.8Hz，1H)，7.64(d，J＝8.4Hz，1H)，8.11(d，J＝9.6Hz，1H)]。

example 31

[ (R) - (2- (N-piperidinyl) ethylthio) methyl-Sar ] -3- [ (gamma-hydroxy) -N-MeLeu ] -4-cyclosporin

Reacting [ alpha-methylene-Sar]-3- [ (γ -hydroxy) -N-MeLeu]-4-Cyclosporin (0.37g, 0.30mmol) and 2- (N-piperidine) ethylthiol (0.44g, 3.00mmol) were dissolved in methanol (30ml), followed by addition of 10 equivalents of lithium hydroxide. The mixture was stirred at room temperature overnight. After removal of the solvent, the residue was dissolved in dichloromethane (30 ml). The dichloromethane solution 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 yield 0.20g of product [ formula: c₇₀H₁₂₆N₁₂O₁₃S; accurate quality: 1374.93, respectively; MS (m/z): 1375.65(M +1)⁺、1397.80(M+Na)⁺；TLC R_f: 0.18 (ethyl acetate/methanol ═ 5/1); HPLC RT: 12.09min. (C8 reverse phase column: 250 mm; acetonitrile/water (0.05% trifluoroacetic acid); operating temperature: 64 ℃ C.; detector: 210nm)]。

Example 32

[ (R) - (2- (4-methyl-N-piperazinyl) ethylthio) methyl-Sar ] -3- [ (gamma-hydroxy) -N-MeLeu ] -4-Cyclosporin

Reacting [ alpha-methylene-Sar]-3- [ (γ -hydroxy) -N-MeLeu]-4-Cyclosporin (0.30g, 0.24mmol) and 2- (4-methylpiperazine) ethylthiol (0.42g, 2.62mmol) were dissolved in methanol (30ml), and 10 equivalents of lithium hydroxide were added. The mixture was stirred at room temperature overnight. After removal of the solvent, the residue was dissolved in dichloromethane (30 ml). The dichloromethane solution 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 yield 0.22g of product [ formula: c₇₀H₁₂₇N₁₃O₁₃S; accurate quality: 1389.94, respectively; MS (m/z): 1390.9(M +1)⁺；TLC R_f: 0.08 (ethyl acetate/methanol ═ 5/1); HPLC RT: 10.07min (C8 reverse phase column: 250 mm; acetonitrile/water (0.05% trifluoroacetic acid); operating temperature: 64 ℃; detector: 210nm)]。

Example 33

[ (R) - (2- (N-pyrrolidinyl) ethylthio) methyl-Sar ] -3- [ (gamma-hydroxy) -N-MeLeu ] -4-Cyclosporin

To [ alpha-methylene-Sar ]]-3- [ (γ -hydroxy) -N-MeLeu]A solution of-4-cyclosporin (280mg, 0.23mmol) and 2- (N-pyrrolidinyl) ethanethiol (280mg, 2.14mmol) in methanol (30ml) was added with lithium hydroxide (114mg, 4.75 mmol). The reaction mixture was stirred at room temperature overnight. Most of the solvent was evaporated under reduced pressure. Dichloromethane (30ml) and water (30ml) were added and the mixture was separated. The organic layer was washed with water and brine, dried over magnesium sulfate and evaporated under reduced pressure. The residue was purified by chromatography (dichloromethane/methanol 96/4) to yield 126mg of product [ formula: c₆₉H₁₂₄N₁₂O₁₃S; accurate quality: 1360.91, respectively; MS (m/z): 1361.80(M +1)⁺；TLC R_f: 0.23 (dichloromethane/methanol 95/5); HPLC RT: 11.59min (C8 reverse phase column: 250 mm; acetonitrile/0.077% ammonium acetate in water; operating temperature: 64 ℃ C.; detector: 210nm)]。

Example 34

[ (R) -alpha- (2-aminoethylthio) methyl-Sar ] -3- [ (gamma-hydroxy) -N-MeLeu ] -4-Cyclosporin

Reacting [ alpha-methylene-Sar]-3- [ (γ -hydroxy) -N-MeLeu]-4-Cyclosporin (0.86g, 0.70mmol) and 2-aminoethanethiol hydrochloride (0.80g, 7.00mmol) were dissolved in methanol (80ml) and 20 equivalents of lithium hydroxide were added. The mixture was stirred at room temperature overnight. After removal of the solvent, the residue was subjected to flash chromatography using dichloromethane/methanol as eluent, yielding 0.60g of the product [ formula: c₆₅H₁₁₈N₁₂O₁₃S; accurate quality: 1306.87, respectively; MS (m/z): 1307.56(M +1)⁺、1329.73(M+Na)⁺、TLC R_f: 0.025 (dichloromethane/methanol ═ 5/1); HPLC RT: 10.97min (C8 reverse phase column: 250 mm; acetonitrile/water (0.05% trifluoroacetic acid); operating temperature: 64 ℃ C.; detector: 210nm)]。

Example 35

[ (R) -alpha- (2- (N-isopropylamino) ethylthio) methyl-Sar ] -3- [ (gamma-hydroxy) -N-MeLeu ] -4-Cyclosporin

The [ (R) -alpha- (2- (amino) ethylthio) methyl-Sar]-3- [ (γ -hydroxy) -N-MeLeu]-4-Cyclosporin (0.31g, 0.25mmol) and acetone (0.40ml) were dissolved in chloroform (30ml), followed by the addition of 2.5 equivalents of tetramethylammonium triacetoxyborohydride in portions with a few drops of acetic acid. The mixture was stirred at room temperature for two hours. The reaction mixture was then washed with saturated sodium bicarbonate solution and brine, dried over magnesium sulfate and evaporated under reduced pressure. The residue was purified by flash chromatography on silica gel using methylene/methanol as eluent to yield 0.25g of pure product [ formula: c₆₈H₁₂₄N₁₂O₁₃S; accurate quality: 1348.91, respectively; MS (m/z): 1349.59(M +1)⁺；TLC R_f: 0.1 (ethyl acetate/methanol ═ 5/1); HPLC RT: 11.97min (C8 reverse phase column: 250 mm; acetonitrile/water (0.05% trifluoroacetic acid); operating temperature: 64 ℃ C.; detector: 210nm)]。

Example 36

[ (R) - (2- (N-Ethyl-N-isopropylamino) ethylthio) methyl-Sar ] -3- [ (gamma-hydroxy) -N-MeLeu ] -4-Cyclosporin

The reaction product of [ (R) - (2- (N-isopropylamino) ethylthio) methyl-Sar]-3- [ (γ -hydroxy) -N-MeLeu]-4-Cyclosporin (49mg, 0.034mmol) and acetaldehyde (100. mu.l, 37% in water) were mixed with chloroform (10ml) and 2.5 equivalents of tetramethylammonium triacetoxyborohydride were added. The mixture was stirred at room temperature for two hours. The reaction mixture was then washed with saturated sodium bicarbonate solution and brine, dried over magnesium sulfate and evaporated under reduced pressure. The residue was purified by flash chromatography on silica gel using dichloromethane/methanol as eluent to yield 37mg of pure product [ formula: c₇₀H₁₂₈N₁₂O₁₃S; accurate quality: 1376.94, respectively; MS (m/z): 1377.84(M +1)⁺；TLC R_f: 0.15 (ethyl acetate/methanol ═ 5/1); HPLC RT: 12.36min (C8 reverse phase column: 250 mm; acetonitrile/water (0.05% trifluoroacetic acid); operating temperature: 64 ℃ C.; detector: 210nm)]。

Example 37

[ (R) - (2- (N-isopropyl-N-methylamino) ethylthio) methyl-Sar ] -3- [ (gamma-hydroxy) -N-MeLeu ] -4-Cyclosporin

The reaction product of [ (R) - (2- (N-isopropylamino) ethylthio) methyl-Sar]-3- [ (γ -hydroxy) -N-MeLeu]-4-Cyclosporin (49mg, 0.034mmol) and formaldehyde (100. mu.l, 37% in water) were mixed with chloroform (10ml) and then 2.5 equivalents of tetramethylammonium triacetoxyborohydride were added. The mixture was stirred at room temperature for two hours. The reaction mixture was then washed with saturated sodium bicarbonate solution and brine, dried over magnesium sulfate and evaporated under reduced pressure. The residue was purified by flash chromatography on silica gel using dichloromethane/methanol as eluent to yield 30mg of pure product [ formula: c₆₉H₁₂₆N₁₂O₁₃S; accurate quality: 1362.93, respectively; MS (m/z): 1363.72(M +1)⁺、1385.81(M+Na)⁺；TLC R_f: 0.15 (ethyl acetate/methanol ═ 5: 1); HPLC RT: 12.26min (C8 reverse phase column: 250 mm; acetonitrile/water (0.05% trifluoroacetic acid); operating temperature: 64 ℃ C.; detector: 210nm)]。

Example 38

[ (R) - (2- (N, N-diisobutylamino) ethylthio) methyl-Sar ] -3- [ (gamma-hydroxy) -N-MeLeu ] -4-Cyclosporin

The [ (R) -alpha- (2-aminoethylthio) methyl-Sar]-3- [ (γ -hydroxy) -N-MeLeu]-4-Cyclosporin (42mg, 0.032mmol) and isobutyraldehyde (15. mu.l) were dissolved in chloroform (10ml), and then 2.5 equivalents of tetramethylammonium triacetoxyborohydride were added in portions. The mixture was stirred at room temperature for two hours. The reaction mixture was then washed with saturated sodium bicarbonate solution and brine, dried over magnesium sulfate and evaporated under reduced pressure. The residue was purified by flash chromatography on silica gel using dichloromethane/methanol as eluent to yield 23mg of pure product [ formula: c₇₃H₁₃₄N₁₂O₁₃S; accurate quality: 1418.99, respectively; MS (m/z): 1419.73(M +1)⁺、1441.87(M+Na)⁺；TLC R_f: 0.36 (ethyl acetate/methanol ═ 5: 1); HPLC RT: 14.46min (C8 reverse phase column: 250 mm; acetonitrile/water (0.05% trifluoroacetic acid); operating temperature: 64 ℃ C.; detector: 210nm)]。

Example 39

[ (R) - (2- (N-isobutylamino-N-isopropyl) ethylthio) methyl-Sar ] -3- [ (γ -hydroxy) -N-MeLeu ] -4-Cyclosporin

The reaction product of [ (R) - (2- (N-isopropylamino) ethylthio) methyl-Sar]-3- [ (γ -hydroxy) -N-MeLeu]-4-Cyclosporin (0.25g, 0.20mmol) and isobutyraldehyde (91. mu.l, 10mmol) were dissolved in chloroform (30ml), and then 2.5 equivalents of tetramethylammonium triacetoxyborohydride were added in portions. The mixture was stirred at room temperature for two hours. The reaction mixture was then washed with saturated sodium bicarbonate solution and brine, dried over magnesium sulfateAnd evaporated under reduced pressure. The residue was purified by flash chromatography on silica gel using dichloromethane/methanol as eluent to yield 19mg of pure product [ formula: c₇₂H₁₃₂N₁₂O₁₃S; accurate quality: 1404.98, respectively; MS (m/z): 1405.89(M +1)⁺、1427.94(M+Na)⁺；TLC R_f: 0.25 (ethyl acetate/methanol ═ 5/1); HPLC RT: 14.46min (C8 reverse phase column: 250 mm; acetonitrile/water (0.05% trifluoroacetic acid); operating temperature: 64 ℃ C.; detector: 210nm)]。

Example 40

[ (R) - (2- (N-neopentylamino) ethylthio) methyl-Sar ] -3- [ (gamma-hydroxy) -N-MeLeu ] -4-Cyclosporin

Reacting [ (R) - (2- (amino) ethylthio) methyl-Sar]-3- [ (γ -hydroxy) -N-MeLeu]-4-Cyclosporin (0.45g, 0.34mmol) and pivalaldehyde (100. mu.l, 37% in water) were mixed with chloroform (50ml) and then 2.5 equivalents of tetramethylammonium triacetoxyborohydride were added. The mixture was stirred at room temperature for two hours. The reaction mixture was then washed with saturated sodium bicarbonate solution and brine, dried over magnesium sulfate and evaporated under reduced pressure. The residue was purified by flash chromatography on silica gel using dichloromethane/methanol as eluent to yield 11mg of pure product [ formula: c₇₀H₁₂₈N₁₂O₁₃S; accurate quality: 1376.94, respectively; MS (m/z): 1377.72(M +1)⁺、1399.82(M+Na)⁺；TLC R_f: 0.15 (ethyl acetate/methanol ═ 5/1); HPLCRT: 12.36min (C8 reverse phase column: 250 mm; acetonitrile/water (0.05% trifluoroacetic acid); operating temperature: 64 ℃ C.; detector: 210nm)]。

Practice ofExample 41

[ (R) - (2- (N-methyl-N-neopentylamino) ethylthio) methyl-Sar ] -3- [ (γ -hydroxy) -N-MeLeu ] -4-Cyclosporin

Mixing [ (R) - (2- (N-neopentyl amino) ethyl sulfur) methyl-Sar]-3- [ (γ -hydroxy) -N-MeLeu]-4-Cyclosporin (49mg, 0.034mmol) and formaldehyde (100. mu.l, 37% in water) were mixed with chloroform (10ml) and then 2.5 equivalents of tetramethylammonium triacetoxyborohydride were added. The mixture was stirred at room temperature for two hours. The reaction mixture was then washed with saturated sodium bicarbonate solution and brine, dried over magnesium sulfate and evaporated under reduced pressure. The residue was purified by flash chromatography on silica gel using dichloromethane/methanol as eluent to yield 31mg of pure product [ formula: c₇₁H₁₃₀N₁₂O₁₃S; accurate quality: 1390.96, respectively; MS (m/z): 1391.71(M +1)⁺、1413.86(M+Na)⁺；TLC R_f: 0.25 (ethyl acetate/methanol ═ 5/1); HPLC RT: 13.28min (C8 reverse phase column: 250 mm; acetonitrile/water (0.05% trifluoroacetic acid); operating temperature: 64 ℃ C.; detector: 210nm)]。

Example 42

[ (R) - (2- (N-Ethyl-N-neopentylamino) ethylthio) methyl-Sar ] -3- [ (gamma-hydroxy) -N-MeLeu ] -4-Cyclosporin

Mixing [ (R) - (2- (N-neopentyl amino) ethyl sulfur) methyl-Sar]-3- [ (γ -hydroxy) -N-MeLeu]-4-Cyclosporin (46mg, 0.034mmol) and acetaldehyde (10. mu.l, 0.17mmol) were dissolved in chloroform/methanolThen 2.5 equivalents of tetramethylammonium triacetoxyborohydride are added in portions. The mixture was stirred at room temperature for two hours. The reaction mixture was then washed with saturated sodium bicarbonate solution and brine, dried over magnesium sulfate and evaporated under reduced pressure. The residue was purified by flash chromatography on silica gel using dichloromethane/methanol as eluent to yield 28mg of product [ formula: c₇₂H₁₃₂N₁₂O₁₃S; accurate quality: 1404.98, respectively; MS (m/z): 1405.75(M +1)⁺、1427.95(M+Na)⁺；TLC R_f: 0.25 (ethyl acetate/methanol ═ 5/1); HPLC RT: 13.65min (C8 reverse phase column: 250 mm; acetonitrile/water (0.05% trifluoroacetic acid); operating temperature: 64 ℃ C.; detector: 210nm)]。

Example 43

[ (R) - (3- (N-piperidinyl) propylthio) methyl-Sar ] -3- [ (gamma-hydroxy) -N-MeLeu ] -4-cyclosporin

To [ alpha-methylene-Sar ]]-3- [ (γ -hydroxy) -N-MeLeu]A solution of-4-cyclosporin (250mg, 0.20mmol) and 3- (N-piperidinyl) propanethiol (318mg, 2.00mmol) in methanol (30ml) was added with lithium hydroxide (96mg, 4.00 mmol). The reaction mixture was stirred at room temperature overnight. Most of the solvent was then evaporated under reduced pressure. Dichloromethane (30ml) and water (30ml) were added and the mixture was separated. The organic layer was washed with water and brine, dried over magnesium sulfate and evaporated under reduced pressure. The residue was purified by chromatography (dichloromethane/methanol 96/4) to yield 135mg of product [ formula: c₇₁H₁₂₈N₁₂O₁₃S; accurate quality: 1388.94, respectively; MS (m/z): 1389.84(M +1)⁺(ii) a TLC Rf: 0.30 (dichloromethane/methanol 95/5); HPLC RT: 12.19min (C8 reverse phase column: 250mm, acetonitrile/0.077% ammonium acetate in water; operating temperature: 64 ℃ C.; detector: 2)10nm)；¹H NMR Spectrum (600MHz, CDCl)₃δ in ppm): 0.68(d, J ═ 6.0Hz, 3H), 0.79(d, J ═ 6.0Hz, 3H), 0.82-0.86(m, 9H), 0.88(d, J ═ 6.6Hz, 3H), 0.91(d, J ═ 6.6Hz, 3H), 0.93(d, J ═ 6.6Hz, 3H), 0.97-1.00(m, 9H), 1.09(d, J ═ 6.6Hz, 3H), 1.21-1.24(m, 11H), 1.31-1.46(m, 8H), 1.53(m, 5H), 1.61(m, 11H), 1.67-1.70(m, 2H), 1.74-1.76(m, 2H), 1.99-2.11(m, 4H), 2.35(m, 2H), 2.35(m, 3.6H, 3H), 3.67-1.70 (m, 2H), 1.74-1.35 (m, 2H), 3.3.48 (m, 3H), 3.48H), 3H) 3.56(m, 1H), 3.65(m, 1H), 4.51(m, 1H), 4.58(t, J ═ 8.4Hz, 1H), 4.81(m, 1H), 4.94-5.02(m, 2H), 5.04(t, J ═ 6.6Hz, 1H), 5.08(d, J ═ 10.8Hz, 1H), 5.28-5.32(m, 1H), 5.33-5.37(m, 1H), 5.49(m, 2H), 5.67(dd, J ═ 10.8Hz and 3.6Hz, 1H), 7.14(d, J ═ 8.4Hz, 1H), 7.49(d, J ═ 8.4Hz, 1H), 7.64(d, J ═ 8.4, 1H), 8.09 (J ═ 8.8, 1H), 10.09 (d, 1H)]。

Example 44

[ (R) - (3- (N-pyrrolidinyl) propylthio) methyl-Sar ] -3- [ (gamma-hydroxy) -N-MeLeu ] -4-Cyclosporin

To [ alpha-methylene-Sar ]]-3- [ (γ -hydroxy) -N-MeLeu]A solution of-4-cyclosporin (213mg, 0.17mmol) and 3- (N-pyrrolidinyl) propanethiol (280mg, 1.93mmol) in methanol (25ml) was added with lithium hydroxide (94mg, 3.92 mmol). The reaction mixture was stirred at room temperature overnight. Most of the solvent was then evaporated under reduced pressure. Dichloromethane (30ml) and water (30ml) were added and the mixture was separated. The organic layer was washed with water and brine, dried over magnesium sulfate and evaporated under reduced pressure. The residue was purified by chromatography (dichloromethane/methanol 95/5) to yield 57mg of product [ formula: c₇₀H₁₂₆N₁₂O₁₃S; accurate quality: 1374.93, respectively; MS (m/z): 1375.75(M +1)⁺(ii) a TLC Rf: 0.23 (dichloromethane/methanol 95/5); HPLC RT: 11.83min (C8 reverse phase column: 250 mm; acetonitrile/0.077% ammonium acetate in water; operating temperature: 64 ℃ C.; detector: 210nm)]。

Example 45

[ (R) - (3- (N-morpholinyl) propylthio) methyl-Sar ] -3- [ (γ -hydroxy) -N-MeLeu ] -4-Cyclosporin

To [ alpha-methylene-Sar ]]-3- [ (γ -hydroxy) -N-MeLeu]A solution of-4-cyclosporin (210mg, 0.17mmol) and 3-morpholinopropanethiol (300mg, 1.86mmol) in methanol 25ml was added lithium hydroxide (140mg, 5.83 mmol). The reaction mixture was stirred at room temperature overnight. Most of the solvent was then evaporated under reduced pressure. Dichloromethane (30ml) and water (30ml) were added and the mixture was separated. The organic layer was washed with water and brine, dried over magnesium sulfate and evaporated under reduced pressure. The residue was purified by chromatography (dichloromethane/methanol 97/3) to yield 78mg of product [ formula: c₇₀H₁₂₆N₁₂O₁₄S; accurate quality: 1390.92, respectively; MS (m/z): 1413.77(M + Na)⁺(ii) a TLC Rf: 0.33 (dichloromethane/methanol ═ 9/1); HPLC RT: 11.35min (C8 reverse phase column: 250 mm; acetonitrile/0.077% ammonium acetate in water; operating temperature: 64 ℃ C.; detector: 210 nm);¹h NMR Spectrum (600MHz, CDCl)₃δ in ppm): 0.68(d, J ═ 6.0Hz, 3H), 0.79(d, J ═ 5.4Hz, 3H), 0.82-0.86(m, 9H), 0.88(d, J ═ 6.6Hz, 3H), 0.91(d, J ═ 6.6Hz, 3H), 0.93(d, J ═ 6.6Hz, 3H), 0.97-1.00(m, 9H), 1.09(d, J ═ 6.6Hz, 3H), 1.21-1.24(m, 11H), 1.31(d, J ═ 7.2Hz, 3H), 1.38-1.46(m, 2H), 1.61(m, 11H), 1.67-1.70(m, 2H), 1.74-1.76(m, 2H), 2.03-2.35 (m, 2H), 4.35-2H, 4.6 (m, 3H), 4H), 3H, 1.6H, 13(m，8H)，2.55-2.63(m，2H)，2.67(s，6H)，2.91-2.98(m.2H)，3.10(s，3H)，3.24(3，6H)，3.26(s，3H)，3.49(s，3H)，3.52(m，1H)，3.65-3.67(m，5H)，4.51(m，1H)，4.59(t，J＝8.4Hz，1H)，4.81(m，1H)，4.94-5.01(m，2H)，5.04(t，J＝6.6Hz，1H)，5.08(d，J＝12Hz，1H)，5.28-5.30(m，1H)，5.33-5.37(m，1H)，5.49(m，2H)，5.67(m，1H)，7.14(d，J＝7.8Hz，1H)，7.49(d，J＝8.4Hz，1H)，7.65(d，J＝7.2Hz，1H)，8.12(d，J＝9.6Hz，1H)]。

Example 46

[ (R) - (3- (4-methyl-N-piperazinyl) propylthio) methyl-Sar ] -3- [ (gamma-hydroxy) -N-MeLeu ] -4-Cyclosporin

Reacting [ alpha-methylene-Sar]-3- [ (γ -hydroxy) -N-MeLeu]-4-Cyclosporin (0.30g, 0.24mmol) and 3- (4-methylpiperazine) propylthiol (0.42g, 2.44mmol) were dissolved in methanol (25ml), and 10 equivalents of lithium hydroxide were added. The mixture was stirred at room temperature overnight. After removal of the solvent, the residue was purified by flash chromatography using dichloromethane/methanol as eluent to yield 0.20g of product [ formula: c₇₁H₁₂₉N₁₃O₁₃S; accurate quality: 1403.96, respectively; MS (m/z): 1404.9(M +1)⁺、1426.9(M+Na)⁺；TLC R_f: 0.10 (ethyl acetate/methanol ═ 5/1); HPLC RT: 10.07min (C8 reverse phase column: 250 mm; acetonitrile/water (0.05% trifluoroacetic acid); operating temperature: 64 ℃; detector: 210nm)]。

Example 47

[ (R) - (3- (N, N-dimethylamino) propylthio) methyl-Sar ] -3- [ (gamma-hydroxy) -N-MeLeu ] -4-Cyclosporin

Reacting [ alpha-methylene-Sar]-3- [ (γ -hydroxy) -N-MeLeu]-4-Cyclosporin (0.30g, 0.24mmol) and 3- (N, N-dimethyl) propyl mercaptan (0.36g, 2.40mmol) were dissolved in methanol (25ml) and lithium hydroxide (59mg, 2.44mmol) was added. The mixture was stirred at room temperature overnight. After removal of the solvent, the residue was purified by flash chromatography using dichloromethane/methanol as eluent to yield 0.18g of pure product [ formula: c₆₈H₁₂₄N₁₂O₁₃S; accurate quality: 1348.91, respectively; MS (m/z): 1349.70(M +1)⁺、1371.83(M+Na)；TLC R_f: 0.15 (ethyl acetate/methanol ═ 5/1); HPLC RT: 11.53min (C8 reverse phase column: 250 mm; acetonitrile/water (0.05% trifluoroacetic acid); operating temperature: 64 ℃ C.; detector: 210nm)]。

Example 48

[ (R) - (3- (N, N-diethylamino) propylthio) methyl-Sar ] -3- [ (gamma-hydroxy) -N-MeLeu ] -4-Cyclosporin

Reacting [ alpha-methylene-Sar]-3- [ (γ -hydroxy) -N-MeLeu]-4-Cyclosporin (0.30g, 0.24mmol) and 3- (N, N-dimethyl) propyl mercaptan (0.36g, 2.44mmol) were dissolved in methanol (25ml) and lithium chloride (59mg, 2.4mmol) was added. The mixture was stirred at room temperature overnight. After removal of the solvent, the residue was purified by flash chromatography using dichloromethane/methanol as eluent to yield 0.30g of the product [ formula: c₇₀H₁₂₈N₁₂O₁₃S; accurate quality: 1376.94, respectively; MS (m/z): 1377.90(M +1))⁺、1399.76(M+Na)⁺；TLC R_f: 0.17 (ethyl acetate/methanol ═ 5/1); HPLC RT: 12.06 min. (C8 reverse phase column: 250 mm; acetonitrile/water (0.05% trifluoroacetic acid); operating temperature: 64 ℃ C.; detector: 210nm)]。

Example 49

[ (R) - (3- (N-Ethyl-N-isopropylamino) propylthio) methyl-Sar ] -3- [ (gamma-hydroxy) -N-MeLeu ] -4-Cyclosporin

To [ alpha-methylene-Sar ]]-3- [ (γ -hydroxy) -N-MeLeu]A solution of-4-cyclosporin (200mg, 0.16mmol) and 3- (N-ethyl-N-isopropylamino) propyl thiol (200mg, 1.25mmol) in methanol (25ml) was added with lithium hydroxide (89mg, 3.71 mmol). The reaction mixture was stirred at room temperature overnight. Most of the solvent was then evaporated under reduced pressure. Dichloromethane (30ml) and water (30ml) were added and the mixture was separated. The organic layer was washed with water and brine, dried over magnesium sulfate and evaporated under reduced pressure. The residue was purified by chromatography (dichloromethane/methanol 97/3) to yield 88mg of product [ formula: c₇₁H₁₃₀N₁₂O₁₃S; accurate quality: 1390.96, respectively; MS (m/z): 1413.81(M + Na)⁺(ii) a TLC Rf: 0.40 (dichloromethane/methanol ═ 9/1); HPLC RT: 12.49min (C8 reverse phase column: 250 mm; acetonitrile/0.077% ammonium acetate in water; operating temperature: 64 ℃ C.; detector: 210nm)]。

Example 50

[ (R) - (2- (N, N-diethylamino) ethoxy) methyl-Sar ] -3- [ (gamma-hydroxy) -N-MeLeu ] -4-Cyclosporin

To [ (R) -alpha-hydroxymethyl-Sar]-3- [ (γ -hydroxy) -N-MeLeu]A solution of-4-cyclosporin (0.25g, 0.20mmol) in benzene (30ml) was added a solution of sodium hydroxide (1.00g, 25mmol) in water (2ml), 2-bromo-N, N-diethylethanamine hydrobromide (2.80g, 10.72mmol) and tetra-N-butylammonium bromide (0.2g, 0.62 mmol). The mixture was stirred at 30 ℃ for 20 hours. Ice water (30ml) was then added and the mixture was separated. The aqueous layer was extracted with dichloromethane (25 ml). The combined organic layers were washed with brine, dried over magnesium sulfate and concentrated under reduced pressure. The residue was purified by chromatography (dichloromethane/methanol 96/4) to yield 240mg of product [ formula: c₆₉H₁₂₆N₁₂O₁₄(ii) a Accurate quality: 1346.95, respectively; MS (m/z): 1347.59(M +1)⁺(ii) a TLC Rf: 0.41 (dichloromethane/methanol ═ 9/1); HPLC RT: 12.20min (C8 reverse phase column: 250 mm; acetonitrile/0.077% ammonium acetate in water; operating temperature: 64 ℃ C.; detector: 210nm)]。

Example 51

[ (R) - (2- (N-piperidinyl) ethoxy) methyl-Sar ] -3- [ (gamma-hydroxy) -N-MeLeu ] -4-cyclosporin

To [ (R) -alpha-hydroxymethyl-Sar]-3- [ (γ -hydroxy) -N-MeLeu]A solution of-4-cyclosporin (300mg, 0.24mmol) in benzene (15ml) was added sodium hydroxide (0.38g.9.60mmol), tetramethylammonium hydroxide pentahydrate (0.44g, 2.40mmol) and 1- (2-chloroethyl) piperidine hydrochloride (0.44g, 2.40 mmol). The mixture was stirred at 30 ℃ for 36 hours. Ice water (20ml) was then added and the mixture was separated. The aqueous layer was extracted with ethyl acetate (20 ml). The combined organic layers were washed with brine, dried over magnesium sulfate andevaporated under reduced pressure. The residue was purified by chromatography on silica gel using dichloromethane/methanol (95/5) as eluent to yield 100mg of pure product [ formula: c₇₀H₁₂₆N₁₂O₁₄(ii) a Accurate quality: 1358.95, respectively; MS (m/z): 1359.69(M +1)⁺、1381.75(M+Na)⁺；TLC R_f: 0.05 (dichloromethane/methanol ═ 20/1); HPLC RT: 12.43min. (C8 reverse phase column: 150 mm; acetonitrile/water (0.05% trifluoroacetic acid); operating temperature: 64 ℃ C.; detector: 210nm)]。

Example 52

[ (R) - (2- (N-morpholinyl) ethoxy) methyl-Sar ] -3- [ (γ -hydroxy) -N-MeLeu ] -4-Cyclosporin

To [ (R) -alpha-hydroxymethyl-Sar]-3- [ (γ -hydroxy) -N-MeLeu]A solution of-4-cyclosporin (0.50g, 0.40mmol) in benzene (20ml) was added sodium hydroxide (0.64g, 16.00mmol), tetramethylammonium hydroxide pentahydrate (0.72g, 4.00mmol) and 4- (2-chloroethyl) morpholine hydrochloride (0.74g, 4.00 mmol). The mixture was stirred at 30 ℃ for one week. Ice water (20ml) was then added and the mixture was separated. The aqueous layer was extracted with ethyl acetate (20 ml). The combined organic layers were washed with brine, dried over magnesium sulfate and evaporated under reduced pressure. The residue was purified by chromatography on silica gel using dichloromethane/methanol (95/5) as eluent to yield 60mg of product [ formula: c₆₉H₁₂₄N₁₂O₁₅(ii) a Accurate quality: 1360.93, respectively; MS (m/z): 1361.63(M +1)⁺、1383.75(M+Na)⁺；TLC R_f: 0.10 (dichloromethane/methanol 5: 1); HPLCRT: 11.49min (C8 reverse phase column: 150 mm; acetonitrile/water (0.05% trifluoroacetic acid); operating temperature: 64 ℃; detector: 210nm)]。

Example 53

[ (R) - (2- (N, N-dimethylamino) ethoxy) methyl-Sar ] -3- [ (gamma-hydroxy) -N-MeLeu ] -4-Cyclosporin

To [ (R) -alpha-hydroxymethyl-Sar]-3- [ (γ -hydroxy) -N-MeLeu]A solution of-4-cyclosporin (0.37g, 0.30mmol) in benzene (15ml) was added sodium hydroxide (0.48g.12.00mmol), tetramethylammonium hydroxide pentahydrate (0.54g, 3.00mmol) and 3-dimethylaminoethyl chloride hydrochloride (0.43g, 3.00 mmol). The mixture was stirred at 30 ℃ for 36 hours. Ice water (20ml) was then added and the mixture was separated. The aqueous layer was extracted with ethyl acetate (20 ml). The combined organic layers were washed with brine, dried over magnesium sulfate and evaporated under reduced pressure. The residue was purified by chromatography on silica gel using dichloromethane/methanol (95/5) as eluent to yield 90mg of pure product [ formula: c₆₇H₁₂₂N₁₂O₁₄(ii) a Accurate quality: 1318.92, respectively; MS (m/z): 1319.70(M +1)⁺、1341.80(M+Na)⁺)；TLC R_f: 0.05 (dichloromethane/methanol 5: 1); HPLC RT: 11.43min (C8 reverse phase column: 150 mm; acetonitrile/water (0.05% trifluoroacetic acid); operating temperature: 64 ℃ C.; detector: 210nm)]。

Example 54

[ (R) - (2- (N-pyrrolidinyl) ethoxy) methyl-Sar ] -3- [ (gamma-hydroxy) -N-MeLeu ] -4-Cyclosporin

To [ (R) -alpha-hydroxymethyl-Sar]-3- [ (γ -hydroxy))-N-MeLeu]A solution of-4-cyclosporin (0.38g, 0.30mmol) in benzene (15ml) was added sodium hydroxide (0.48g, 12.00mmol), tetramethylammonium hydroxide pentahydrate (0.54g, 3.00mmol) and 1- (2-chloroethyl) pyrrolidine hydrochloride (0.44g, 3.00 mmol). The mixture was stirred at 30C for 36 hours. Ice water (20ml) was then added and the mixture was separated. The aqueous layer was extracted with ethyl acetate (20 ml). The combined organic layers were washed with brine, dried over magnesium sulfate and evaporated under reduced pressure. The residue was purified by chromatography on silica gel using dichloromethane/methanol (95/5) as eluent to yield 120mg of the expected isomer [ formula: c₆₉H₁₂₄N₁₂O₁₄(ii) a Accurate quality: 1344.94, respectively; MS (m/z): 1345.62(M +1)⁺、1367.76(M+Na)⁺；TLC R_f: 0.05 (dichloromethane/methanol ═ 10/1); HPLC RT: 12.09min. (C8 reverse phase column: 150 mm; acetonitrile/water (0.05% trifluoroacetic acid); operating temperature: 64 ℃ C.; detector: 210nm)]。

Example 55

[ (R) -alpha- (2- (1, 3-dioxan-2-yl) ethoxy) methyl-Sar ] -3- [ (gamma-hydroxy) -N-MeLeu ] -4-Cyclosporin

Reacting [ (R) -alpha-hydroxymethyl-Sar]-3- [ (γ -hydroxy) -N-MeLeu]-4-Cyclosporin (5.00g, 4.01mmol) was dissolved in benzene (100 ml). 2- (2-bromoethyl) -1, 3-dioxane (7.82g, 40.10mmol), tetra-n-butylammonium bromide (0.99g, 3.09mmol), sodium hydroxide (3.21g, 8.02mmol) and water (3.3ml) were added. The reaction mixture was stirred at 35 ℃ for 9 hours. And stirring was continued overnight at room temperature. Then 50ml of brine were added and the mixture was separated. The aqueous layer was extracted with ethyl acetate (25 ml. times.2). The combined organic layers were dried over magnesium sulfate and evaporated under reduced pressure. After purification on silica gel using hexane/acetone as eluent, 1.50g of product [ formula：C₆₉H₁₂₃N₁₁O₁₆(ii) a Accurate quality: 1361.91, respectively; (m/z): 1362.64(M +1)⁺、1384.85(M+Na)⁺]。

Example 56

[ (R) -alpha- (2-formylethoxy) methyl-Sar ] -3- [ (gamma-hydroxy) -N-MeLeu ] -4-cyclosporin

Reacting [ (R) -alpha- (2- (1, 3-dioxane-2-yl) ethoxy) methyl-Sar]-3- [ (γ -hydroxy) -N-MeLeu]-4-cyclosporin (1.29g, 0.95mmol) was dissolved in dioxane (25ml) and hydrochloric acid solution (1N, 25ml) was added. The reaction mixture was stirred at room temperature overnight. Most of the dioxane was evaporated under reduced pressure. The aqueous layer was then extracted with ethyl acetate (25 ml. times.2). The combined ethyl acetate layers were dried over magnesium sulfate and evaporated under reduced pressure. The residue was purified on silica gel using hexane/acetone as eluent to yield 600mg of product [ formula: c₆₆H₁₁₇N₁₁O₁₅(ii) a Accurate quality: 1303.87, respectively; MS (m/z): 1304.59(M +1)⁺、1326.78(M+Na)⁺(ii) a HPLC RT: 14.2min (C8 reverse phase column: 250 mm; acetonitrile/water (0.05% trifluoroacetic acid); operating temperature: 64 ℃ C.; detector: 210nm)]。

Example 57

[ (R) - (3- (N-morpholinyl) propoxy) methyl-Sar ] -3- [ (γ -hydroxy) -N-MeLeu ] -4-Cyclosporin

Mixing [ (R) - (2-formylethoxy) methyl-Sar]-3- [ (γ -hydroxy) -N-MeLeu]-4-Cyclosporin (300mg, 0.23mmol) was dissolved in dichloromethane (15 ml). Morpholine (100mg, 1.15mmol) and tetramethylammonium triacetoxyborohydride (302mg, 1.15mmol) were added. The reaction mixture was stirred at room temperature overnight. Then a saturated solution of sodium bicarbonate (30ml) and dichloromethane (15ml) were added and the mixture was separated. The dichloromethane layer was dried over magnesium sulfate and evaporated under reduced pressure. After purification on silica gel, 105mg of pure product are obtained [ formula: c₇₀H₁₂₆N₁₂O₁₅(ii) a Accurate quality: 1374.95, respectively; MS (m/z): 1375.70(M +1)⁺、1397.80(M+Na)⁺；TLC R_f: 0.37 (dichloromethane/methanol ═ 9/1); HPLC RT: 12.2min (C8 reverse phase column: 250 mm; acetonitrile/0.077% ammonium acetate in water; operating temperature: 64 ℃ C.; detector: 210nm)]。

Example 58

[ (R) - (3- (N-pyrrolidinyl) propoxy) methyl-Sar ] -3- [ (γ -hydroxy) -N-MeLeu ] -4-Cyclosporin

The [ (R) -alpha- (2-formylethoxy) methyl-Sar]-3- [ (γ -hydroxy) -N-MeLeu]-4-Cyclosporin (200mg, 0.15mmol) was dissolved in dichloromethane (15 ml). Pyrrolidine (95mg, 1.34mmol) and tetramethylammonium triacetoxyborohydride (353mg, 1.34mmol) were added. The reaction mixture was stirred at room temperature overnight. Then a saturated solution of sodium bicarbonate (30ml) and dichloromethane (15ml) were added and the mixture was separated. The dichloromethane layer was dried over magnesium sulfate and evaporated under reduced pressure. After purification on silica gel, 50mg of pure product are obtained [ formula: c₇₀H₁₂₆N₁₂O₁₄(ii) a Accurate quality: 1358.95, respectively; MS (m/z): 1359.74(M +1)⁺、1381.79(M+Na)⁺；TLC R_f：0.40(dichloromethane/methanol ═ 9/1); HPLC RT: 12.7min (C8 reverse phase column: 250 mm; acetonitrile/0.077% ammonium acetate in water; operating temperature: 64 ℃ C.; detector: 210nm)]。

Example 59

[ (R) - (3- (N-piperidinyl) propoxy) methyl-Sar ] -3- [ (gamma-hydroxy) -N-MeLeu ] -4-cyclosporin

The [ (R) -alpha- (2-formylethoxy) methyl-Sar]-3- [ (γ -hydroxy) -N-MeLeu]-4-Cyclosporin (200mg, 0.15mmol) was dissolved in dichloromethane (15 ml). Piperidine (114mg, 1.34mmol) and tetramethylammonium triacetoxyborohydride (353mg, 1.34mmol) were added. The reaction mixture was stirred at room temperature overnight. Then a saturated solution of sodium bicarbonate (30ml) and dichloromethane (15ml) were added and the mixture was separated. The dichloromethane layer was dried over magnesium sulfate and evaporated under reduced pressure. After purification on silica gel, 43mg of product are obtained [ formula: c₇₁H₁₂₈N₁₂O₁₄(ii) a Accurate quality: 1372.97, respectively; (m/z): MS (m/z): 1373.79(M +1)⁺、1395.86(M+Na)⁺；TLC R_f: 0.27 (dichloromethane/methanol ═ 9/1); HPLC RT: 17.8min (C8 reverse phase column: 250 mm; acetonitrile/0.077% ammonium acetate in water; operating temperature: 64 ℃ C.; detector: 210nm)]。

Example 60

[ (R) - (3- (N, N-dimethylamino) propoxy) methyl-Sar ] -3- [ (gamma-hydroxy) -N-MeLeu ] -4-Cyclosporin

To [ (R) -alpha-hydroxymethyl-Sar]-3- [ (γ -hydroxy) -N-MeLeu]A solution of-4-cyclosporin (0.38g, 0.30mmol) in benzene (15ml) was added sodium hydroxide (0.48g, 12.00mmol), tetramethylammonium hydroxide (0.54g, 3.0mmol) and 3-dimethylaminoethyl chloride hydrochloride (0.43g, 3.00 mmol). The mixture was stirred at 30 ℃ for 36 hours. Ice water (20ml) was then added and the mixture was separated. The aqueous layer was extracted with ethyl acetate (20 ml). The combined organic layers were washed with brine, dried over magnesium sulfate and evaporated under reduced pressure. The residue was purified by chromatography on silica gel using dichloromethane/methanol (95/5) as eluent to yield 70mg of pure product [ formula: c₆₈H₁₂₄N₁₂O₁₄(ii) a Accurate quality: MS (m/z): 1333.64(M +1)⁾+、1355.73(M+Na⁾+；TLC R_f: 0.04 (dichloromethane/methanol ═ 5/1); HPLC RT: 11.78min (C8 reverse phase column: 150 mm; acetonitrile/water (0.05% trifluoroacetic acid); operating temperature: 64 ℃ C.; detector: 210nm)]。

Example 61

[ (R) - (3- (N, N-diethylamino) propoxy) methyl-Sar ] -3- [ (gamma-hydroxy) -N-MeLeu ] -4-cyclosporin

To [ (R) -alpha-hydroxymethyl-Sar]-3- [ (γ -hydroxy) -N-MeLeu]A solution of-4-cyclosporin (250mg, 0.20mmol) in benzene (15ml) was added a solution of sodium hydroxide (400mg, 10.00mmol) in water (0.5ml), 3-diethylaminopropyl chloride hydrochloride (500mg, 2.69mmol) and tetramethylammonium hydroxide pentahydrate (430mg, 2.41 mmol). The mixture was stirred at 32 ℃ for 4 days. Ice water (30ml) was then added and the mixture was separated. The aqueous layer was extracted with dichloromethane (50 ml). The combined organic layers were washed with brine, dried over magnesium sulfate and reduced pressureAnd (5) concentrating. The residue was purified by chromatography (dichloromethane/methanol 95/5) to yield 120mg of product [ formula: c₇₀H₁₂₈N₁₂O₁₄(ii) a Accurate quality: 1360.97, respectively; MS (m/z): 1361.72(M +1)⁺(ii) a TLC Rf: 0.38 (dichloromethane/methanol ═ 9/1); HPLC RT: 16.71min (C8 reverse phase column: 250 mm; acetonitrile/0.077% ammonium acetate in water; operating temperature: 64 ℃ C.; detector: 210nm)]。

Example 62

[ (gamma- (methylthio) methoxy) -N-MeLeu ] -4-cyclosporin

To [ (gamma-hydroxy) -N-MeLeu]A solution of-4-cyclosporin (4.50g, 3.70mmol) in anhydrous dimethyl sulfoxide (25ml) was added to acetic anhydride (15 ml). The reaction mixture was stirred at room temperature for 17 hours. After dilution with ethyl acetate (75ml), the mixture was washed with saturated aqueous sodium bicarbonate and brine, dried over magnesium sulfate and evaporated under reduced pressure. The residue was purified by chromatography on silica gel using dichloromethane/methanol (98/2) as eluent to give 2.35g of [ (gamma-methylthio) methoxy-N-MeLeu [ (. gamma. -methylthio) methoxy- ] -N-MeLeu []-4-cyclosporin [ formula: c₆₄H₁₁₅N₁₁O₁₃S; accurate quality: 1277.84, respectively; MS (m/z): 1300.70(M + Na)⁺；TLC R_f: 0.30 (dichloromethane/methanol 95/5); HPLC RT: 19.57min (C8 reverse phase column: 250 mm; acetonitrile/0.077% ammonium acetate in water; operating temperature: 64 ℃ C.; detector: 210nm)]。

Example 63

[ gamma- (methoxy) -N-MeLeu ] -4-Cyclosporin

To [ gamma- (methylthio) methoxy-N-MeLeu ]]A solution of-4-cyclosporin (1.20g, 0.94mmol) in dry tetrahydrofuran (40ml) was added Raney Ni (. about.2 g). The resulting suspension was stirred and heated to 60 ℃ for 30 minutes and the reaction was monitored by LC-MS. The reaction mixture was filtered and the filter cake was washed with tetrahydrofuran. The filtrate was collected and evaporated under reduced pressure. The residue was purified by chromatography using an eluent of ethyl acetate/methanol (97.5/2.5) to yield 0.60g of product [ formula: c₆₃H₁₁₃N₁₁O₁₃(ii) a Accurate quality: 1231.85, respectively; MS (m/z): 1232.70(M +1)⁺；TLC R_f: 0.46 (dichloromethane/methanol 95/5); HPLCRT: 20.63min (C8 reverse phase column: 250 mm; acetonitrile/0.077% ammonium acetate in water; operating temperature: 64 ℃ C.; detector: 210nm)]。

Example 64

[ alpha-carboxy-Sar ] -3- [ (gamma-methoxy) -N-MeLeu ] -4-Cyclosporin

n-BuLi (2.8M in tetrahydrofuran/hexane, 5.00ml, 14.00mmol) was added to a solution of diisopropylamine (1.44g, 14.30mmol) in tetrahydrofuran (30ml) at-78 ℃ under a nitrogen atmosphere. After stirring the mixture for one and a half hours, [ gamma- (methoxy) -N-MeLeu was added slowly]-a solution of 4-cyclosporin (1.20g, 0.97mmol) in tetrahydrofuran (6 ml). Stirring was continued for 2 hours at-78 ℃. Carbon dioxide gas was then bubbled through the reaction mixture for one hour. The mixture was allowed to warm slowly to room temperature and stirred for another 3 hours. After evaporating most of the solvent under reduced pressure, dichloromethane (30ml) and water (30ml) were added. By adding citric acid waterThe solution adjusted the PH of the mixture to around 5. The mixture was separated and the dichloromethane layer was washed with brine, dried over magnesium sulfate and concentrated under reduced pressure to yield 1.20g of crude product for the next step [ formula: c₆₄H₁₁₃N₁₁O₁₅(ii) a Accurate quality: 1275.84, respectively; MS (m/z): 1298.53(M + Na)⁺]。

Example 65

[ alpha-methoxycarbonyl-Sar ] -3- [ (gamma-methoxy) -N-MeLeu ] -4-cyclosporin

To [ alpha-carboxyl group]-3- [ (γ -methoxy) -N-MeLeu]A mixture of-4-cyclosporin (1.20g.0.94mmol) and potassium carbonate (0.80g, 5.79mmol) in N, N-dimethylformamide (25ml) was added methyl iodide (0.80g, 5.63 mmol). The mixture was stirred at room temperature overnight. Dichloromethane (75ml) and water (30ml) were added and the mixture was separated. The dichloromethane layer was washed with water (25ml) and brine (25ml), dried over magnesium sulfate and concentrated under reduced pressure to give 1.10g of crude product [ formula: c₆₅H₁₁₅N₁₁O₁₅(ii) a Accurate quality: 1289.86, respectively; MS (m/z): 1312.72(M + Na)⁺]。

Example 66

[ (R) -alpha-hydroxymethyl-Sar ] -3- [ (gamma-methoxy) -N-MeLeu ] -4-cyclosporin

To [ alpha-methoxycarbonyl-Sar ]]-3- [ (γ -methoxy) -N-MeLeu]-4-Cyclosporium spA suspension of a bacteriocin (1.10g, 0.85mmol) and lithium chloride (1.00g, 23.53mmol) in methanol (80ml) was added portionwise to sodium borohydride (2.00g, 52.91 mmol). The mixture was stirred at room temperature overnight and concentrated under reduced pressure. Dichloromethane (50ml) and water (30ml) 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 97/3) to yield 310mg of product [ formula: c₆₄H₁₁₅N₁₁O₁₄(ii) a Accurate quality: 1261.86, respectively; MS (m/z): 1262.68(M +1)⁺]。

Example 67

[ alpha-methylene-Sar ] -3- [ (gamma-methoxy) -N-MeLeu ] -4-Cyclosporin

[ alpha-methylene-Sar ]]-3- [ (γ -methoxy) -N-MeLeu]-4-cyclosporin was prepared according to the method described in examples 4 and 5 [ formula: c₆₄H₁₁₃N₁₁O₁₃(ii) a Accurate quality: 1243.85, respectively; MS (m/z): 1244.57(M +1)⁺；TLC R_f: 0.34 (hexane/acetone ═ 6/1); HPLC RT: 17.10min (C8 reverse phase column: 250 mm; acetonitrile/water (0.05% trifluoroacetic acid), operating temperature: 64 ℃ and detector: 210nm]。

Example 68

[ (R) - (2- (N, N-dimethylamino) ethylthio) methyl-Sar ] -3- [ (gamma-methoxy) -N-MeLeu ] -4-Cyclosporin

Reacting [ alpha-methylene-Sar]-3- [ (γ -methoxy) -N-MeLeu]-4-Cyclosporin (0.24g, 0.19mmol) and 2- (N, N-dimethylamino) ethylthiol hydrochloride (0.27g, 1.91mmol) were dissolved in methanol (30ml) and 20 equivalents of lithium hydroxide were added. The mixture was stirred at room temperature overnight. After removal of the solvent, the residue was purified by flash chromatography using dichloromethane/methanol (96/4) as eluent to yield 0.11g of pure product [ formula: c₆₈H₁₂₄N₁₂O₁₃S; accurate quality: 1348.93, respectively; MS (m/e): 1349.85(M +1)⁺、1371.81(M+Na)⁺；TLC R_f: 0.20 (ethyl acetate/methanol (5: 1); HPLC RT: 12.42min. (C8 reverse phase column: 250 mm; acetonitrile/water (0.05% trifluoroacetic acid); operating temperature: 64 ℃ C.; detector: 210nm)]。

Example 69

[ (R) - (2- (N, N-diethylamino) ethylthio) methyl-Sar ] -3- [ (gamma-methoxy) -N-MeLeu ] -4-Cyclosporin

To [ alpha-methylene-Sar ]]-3- [ (γ -methoxy) -N-MeLeu]A solution of-4-cyclosporin (280mg, 0.23mmol) and 2-diethylaminoethanethiol hydrochloride (570mg, 3.37mmol) in methanol (15ml) was added with lithium hydroxide (142mg, 5.92 mmol). The reaction mixture was stirred at room temperature overnight. Most of the solvent was evaporated under reduced pressure. Dichloromethane (80ml) and water (30ml) were added and the mixture was separated. The organic layer was washed with water and brine, dried over magnesium sulfate and evaporated under reduced pressure. The residue was purified by chromatography (dichloromethane/methanol 97/3) to yield 110mg of product [ formula: c₇₀H₁₂₈N₁₂O₁₃S; accurate quality: 1376.94, respectively; MS (m/z): 1377.67(M +1)⁺；TLC R_f: 0.35 (methylene chloride/methylene chloride)Alcohol 95/5); HPLCRT: 13.17min (C8 reverse phase column: 250 mm; acetonitrile/water (0.05% trifluoroacetic acid); operating temperature: 64 ℃ C.; detector: 210nm)]。

Example 70

[ (R) - (2- (N-pyrrolidinyl) ethylthio) methyl-Sar ] -3- [ (gamma-methoxy) -N-MeLeu ] -4-Cyclosporin

To [ alpha-methylene-Sar ]]-3- [ (γ -methoxy) -N-MeLeu]A solution of-4-cyclosporin (230mg, 0.18mmol) and 2- (N-pyrrolidinyl) ethanethiol (340mg, 2.59mmol) in methanol (15ml) was added with lithium hydroxide (120mg, 5.00 mmol). The reaction mixture was stirred at room temperature overnight. Most of the solvent was evaporated under reduced pressure. Dichloromethane (30ml) and water (30ml) were added and the mixture was separated. The organic layer was washed with water and brine, dried over magnesium sulfate and evaporated under reduced pressure. The residue was purified by chromatography (dichloromethane/methanol 96/4) to yield 55mg of product [ formula: c₇₀H₁₂₆N₁₂O₁₃S; accurate quality: 1374.93, respectively; MS (m/z): 1375.57(M +1)⁺；TLC R_f: 0.29 (dichloromethane/methanol 95/5); HPLC RT: 12.90min (C8 reverse phase column: 250 mm; acetonitrile/water (0.05% trifluoroacetic acid); operating temperature: 64 ℃ C.; detector: 210nm)]。

Example 71

[ (R) - (2- (N-morpholinyl) ethylthio) methyl-Sar ] -3- [ (gamma-methoxy) -N-MeLeu ] -4-Cyclosporin

To [ alpha-methylene-Sar ]]-3- [ (γ -methoxy) -N-MeLeu]A solution of-4-cyclosporin (250mg, 0.20mmol) and 2-morpholinoethanethiol (260mg, 1.76mmol) in methanol (15ml) was added with lithium hydroxide (120mg, 5.00 mmol). The reaction mixture was stirred at room temperature overnight. Most of the solvent was evaporated under reduced pressure. Dichloromethane (60ml) and water (30ml) were added and the mixture was separated. The organic layer was washed with water and brine, dried over magnesium sulfate and evaporated under reduced pressure. The residue was purified by chromatography (dichloromethane/methanol 97/3) to yield 70mg of product [ formula: c₇₀H₁₂₆N₁₂O₁₄S; accurate quality: 1390.92, respectively; MS (m/z): 1391.58(M +1)⁺(ii) a TLC Rf: 0.38 (dichloromethane/methanol ═ 9/1); HPLC RT: 12.48min (C8 reverse phase column: 250 mm; acetonitrile/water (0.05% trifluoroacetic acid); operating temperature: 64 ℃ C.; detector: 210nm)]。

Example 72

[ (R) - (3- (N, N-dimethylamino) propylthio) methyl-Sar ] -3- [ (gamma-methoxy) -N-MeLeu ] -4-Cyclosporin

Reacting [ alpha-methylene-Sar]-3- [ (γ -methoxy) -N-MeLeu]-4-Cyclosporin (0.25g, 0.20mmol) and 3- (N, N-dimethylamino) propyl mercaptan (0.24g, 2.00mmol) were dissolved in methanol (20ml) and 10 equivalents of lithium hydroxide were added. The mixture was stirred at room temperature overnight. After removal of the solvent, the residue was purified by flash chromatography using dichloromethane/methanol as eluent to yield 80mg of pure product [ formula: c₆₉H₁₂₆N₁₂O₁₃S; accurate quality: 1362.93, respectively; MS (m/e): 1363.70(M +1)⁺.TLCR_f: 0.20 (ethyl acetate/methanol ═ 10/1); HPLC RT: 12.82min. (C8 reverse phase column: 250 mm; acetonitrile/water (0.05%)Trifluoroacetic acid); operating temperature: 64 ℃; a detector: 210nm)]。

Example 73

[ (R) - (3- (N, N-diethylamino) propylthio) methyl-Sar ] -3- [ (gamma-methoxy) -N-MeLeu ] -4-Cyclosporin

Reacting [ alpha-methylene-Sar]-3- [ (γ -methoxy) -N-MeLeu]-4-Cyclosporin (0.25g, 0.20mmol) and 3- (N, N-diethylamino) propyl mercaptan (0.30g, 2.00mmol) were dissolved in methanol (20ml), and 10 equivalents of lithium hydroxide were added. The mixture was stirred at room temperature overnight. After removal of the solvent, the residue was purified by flash chromatography using dichloromethane/methanol as eluent to yield 120mg of pure product [ formula: c₇₁H₁₃₀N₁₂S; accurate quality: 1390.96, respectively; MS (m/e): 1391.64(M +1)⁺、1413.79(M+Na)⁺；TLC R_f: 0.25 (ethyl acetate/methanol ═ 10/1); HPLC RT: 13.57min (C8 reverse phase column: 250 mm; acetonitrile/water (0.05% trifluoroacetic acid); operating temperature: 64 ℃ C.; detector: 210nm)]。

Example 74

[ (R) - (3- (N-piperidinyl) propylthio) methyl-Sar ] -3- [ (gamma-methoxy) -N-MeLeu ] -4-cyclosporin

Reacting [ alpha-methylene-Sar]-3- [ (γ -methoxy) -N-MeLeu]-4-cyclosporin (0.37g, 0.30mmol) and 3- (N-piperidine) propyl thiol (0.48g,3.00mmol) was dissolved in methanol (30ml) and 10 equivalents of lithium hydroxide were added. The mixture was stirred at room temperature overnight. After removal of the solvent, the residue was purified by flash chromatography using dichloromethane/methanol as eluent to yield 60mg of pure product [ formula: c₇₂H₁₃₀N₁₂O₁₃S; accurate quality: 1402.96, respectively; MS (m/e): 1403.69(M +1)⁺、1425(M+Na)⁺；TLC：R_f: 0.3 (ethyl acetate/methanol ═ 10/1); HPLC RT: 13.59min (C8 reverse phase column: 250 mm; acetonitrile/water (0.05% trifluoroacetic acid); operating temperature: 64 ℃ C.; detector: 210nm)]。

Example 75

[ (R) - (3- (N-pyrrolidinyl) propylthio) methyl-Sar ] -3- [ (gamma-methoxy) -N-MeLeu ] -4-Cyclosporin

To [ alpha-methylene-Sar ]]-3- [ (γ -methoxy) -N-MeLeu]A solution of-4-cyclosporin (280mg, 0.23mmol) and 3- (N-pyrrolidinyl) propanethiol (350mg, 2.41mmol) in methanol (15ml) was added with lithium hydroxide (120mg, 5.00 mmol). The reaction mixture was stirred at room temperature overnight. Most of the solvent was then evaporated under reduced pressure. Dichloromethane (80ml) and water (25ml) were added and the mixture was separated. The organic layer was washed with water and brine, dried over magnesium sulfate and evaporated under reduced pressure. The residue was purified by chromatography (dichloromethane/methanol 94/6) to yield 47mg of product [ formula: c₇₁H₁₂₈N₁₂O₁₃S; accurate quality: 1388.94, respectively; MS (m/z): 1389.68(M +1)⁺(ii) a TLC Rf: 0.30 (dichloromethane/methanol 95/5); HPLCRT: 13.25min (C8 reverse phase column: 250 mm; acetonitrile/water (0.05% trifluoroacetic acid); operating temperature: 64 ℃ C.; detector: 210nm)]。

Example 76

[ (R) - (3- (N-morpholinyl) propylthio) methyl-Sar ] -3- [ (gamma-methoxy) -N-MeLeu ] -4-Cyclosporin

To [ alpha-methylene-Sar ]]-3- [ (γ -methoxy) -N-MeLeu]A solution of-4-cyclosporin (320mg, 0.26mmol) and 3-morpholinopropanethiol (600mg, 3.73mmol) in methanol (25ml) was added lithium hydroxide (140mg, 5.83 mmol). The reaction mixture was stirred at room temperature overnight. Most of the solvent was then evaporated under reduced pressure. Dichloromethane (60ml) and water (25ml) were added and the mixture was separated. The organic layer was washed with water and brine, dried over magnesium sulfate and evaporated under reduced pressure. The residue was purified by chromatography (dichloromethane/methanol 97/3) to yield 58mg of product [ formula: c₇₁H₁₂₈N₁₂O₁₄S; accurate quality: 1404.94, respectively; MS (m/z): 1405.52(M +1)⁺(ii) a TLC Rf: 0.39 (dichloromethane/methanol ═ 9/1); HPLC RT: 15.96min (C8 reverse phase column: 250 mm; acetonitrile/0.077% ammonium acetate in water; operating temperature: 64 ℃ C.; detector: 210nm)]。

Example 77

[ (R) - (2- (N, N-diethylamino) ethoxy) methyl-Sar ] -3- [ (gamma-methoxy) -N-MeLeu ] -4-Cyclosporin

To [ (R) -alpha-hydroxymethyl-Sar]-3- [ (γ -methoxy) -N-MeLeu]A solution of-4-cyclosporin (0.20g, 0.16mmol) in benzene (10ml) was added sodium hydroxide (0.48g, 12.00mmol) in water (1ml)Liquid, 2-bromo-N, N-diethylethylamine hydrobromide (1.10g, 4.21mmol) and tetra-N-butylammonium bromide (0.10g, 0.31 mmol). The mixture was stirred at 35 ℃ for 40 hours. Ice water (10ml) was added and the mixture was separated. The aqueous layer was extracted with dichloromethane (20 ml). The combined organic layers were washed with brine, dried over magnesium sulfate and concentrated under reduced pressure. The residue was purified by chromatography (dichloromethane/methanol 96/4) to yield 36mg of product [ formula: c₇₀H₁₂₈N₁₂O₁₄(ii) a Accurate quality: 1360.97, respectively; MS (m/z): 1383.74(M + Na)⁺；TLC R_f: 0.32 (dichloromethane/methanol 95/5); HPLC RT: 13.66min (C8 reverse phase column: 250 mm; acetonitrile/0.077% ammonium acetate in water; operating temperature: 64 ℃ C.; detector: 210 nm);¹h NMR Spectrum (600MHz, CDCl)₃δ in ppm): 0.65(d, J ═ 4.8Hz, 3H), 0.82(m, 9H), 0.87(m, 6H), 0.91(d, J ═ 6Hz, 3H), 0.93(d, J ═ 6.0Hz, 3H), 0.98-1.01(m, 15H), 1.07(d, J ═ 6.6Hz, 3H), 1.11(s, 6H), 1.23(m, 6H), 1.33(d, J ═ 7.2Hz, 3H), 1.39-1.47(m, 2H), 1.53-1.58(m, 4H), 1.6(m, 3H), 1.67-1.75(m, 3H), 1.98-2.12(m, 4H), 2.43-2.48(m, 3H), 2.50-2.54(m, 4H), 2.50 (m, 3H), 3.67-3H, 3.48(m, 4H), 3.52-3.56(m, 1H), 3.60-3.62(m, 1H), 3.67-3.70(m, 1H), 3.80(m, 1H), 4.06(t, J ═ 9.6Hz, 1H), 4.52(m, 1H), 4.57(m, 1H), 4.80(m, 1H), 4.91(t, J ═ 7.8Hz, 1H), 5.04(m, 3H), 5.11(d, J ═ 11.4Hz, 1H), 5.28-5.34(m, 2H), 5.50(d, J ═ 7.2Hz, 1H), 5.67(m, 1H), 7.10(d, J ═ 7.8Hz 1H), 7.48(d, J ═ 7.80, 7.7H, 7.58 (J ═ 7.7H), 7.58(d, 1H), 7.10(d, 1H), 1H), 7.7.7.10 (d, 1H)]。

Example 78

[ (R) - (2- (N, N-dimethylamino) ethoxy) methyl-Sar ] -3- [ (gamma-methoxy) -N-MeLeu ] -4-Cyclosporin

To [ (R) -alpha-hydroxymethyl-Sar]-3- [ (γ -methoxy) -N-MeLeu]A solution of-4-cyclosporin (250mg, 0.20mmol) in benzene (20ml) was added a solution of sodium hydroxide (633mg, 15.85mmol) in water (0.70ml), tetramethylammonium hydroxide pentahydrate (720mg, 3.96mmol) and 2-dimethylaminoethyl chloride hydrochloride (570mg, 3.96 mmol). The mixture was stirred at 40 to 50 ℃ for two days. A solution of sodium hydroxide (633mg, 15.85mmol) in water (0.70ml), tetramethylammonium hydroxide pentahydrate (720mg, 3.96mmol) and 2-dimethylaminoethyl chloride hydrochloride (570mg, 3.96mmol) were added and the mixture was kept under stirring at 40 to 50 ℃ for another two days. Another portion of 2-dimethylaminoethyl chloride hydrochloride (1.14g, 7.91mmol) was added and stirring continued for another day at 40 to 50 ℃. A saturated solution of sodium bicarbonate (30ml) was added and the mixture was separated. The aqueous layer was then extracted with ethyl acetate (25 ml. times.2). The combined organic layers were dried over magnesium sulfate and evaporated under reduced pressure. The residue was dissolved in ethyl acetate (25 ml). The ethyl acetate phase obtained is washed with an acetic acid solution (5ml in 10ml of water) and a saturated solution of sodium bicarbonate (30 ml). Dried over magnesium sulfate and evaporated under reduced pressure. After purification on silica gel, 21mg of product [ formula: c₆₈H₁₂₄N₁₂O₁₄(ii) a Accurate quality: 1332.94, respectively; MS (m/z): 1333.75(M +1)⁺、1355.87(M+Na)⁺；TLC R_f: 0.22 (dichloromethane/methanol ═ 9/1); HPLC RT: 17.3min (C8 reverse phase column: 250 mm; acetonitrile/0.077% ammonium acetate in water; operating temperature: 64 ℃ C.; detector: 210nm)]

Example 79

[ (R) - (2- (N-morpholinyl) ethoxy) methyl-Sar ] -3- [ (gamma-methoxy) -N-MeLeu ] -4-cyclosporin

To [ (R) -alpha-hydroxymethyl-Sar]-3- [ (γ -methoxy) -N-MeLeu]A solution of-4-cyclosporin (250mg, 0.20mmol) in benzene (20ml) was added a solution of sodium hydroxide (633mg, 15.85mmol) in water (0.70ml), followed by tetramethylammonium hydroxide pentahydrate (720mg, 3.96mmol) and 2- (4-morpholinyl) ethyl chloride hydrochloride (737mg, 3.96 mmol). The mixture was stirred at 40 to 50 ℃ for two days. A solution of sodium hydroxide (633mg, 15.85mmol) in water (0.70ml), tetramethylammonium hydroxide pentahydrate (720mg, 3.96mmol) and 2- (4-morpholinyl) ethyl chloride hydrochloride (737mg, 3.96mmol) were added and the mixture was kept under stirring for a further two days at 40 to 50 ℃. Another portion of 2- (4-morpholinyl) ethyl chloride hydrochloride (1.47g, 7.91mmol) was added and stirring was continued for another two days at 40 to 50 ℃. A saturated solution of sodium bicarbonate (30ml) was added and the mixture was separated. The aqueous layer was then extracted with ethyl acetate (25 ml. times.2). The combined organic layers were dried over magnesium sulfate and evaporated under reduced pressure. The residue was dissolved in ethyl acetate (25ml) and the ethyl acetate phase obtained was washed with acetic acid solution (5ml in 10ml water) and saturated solution of sodium bicarbonate (30 ml). Dried over magnesium sulfate and evaporated under reduced pressure. After purification on silica gel, 45mg of product are obtained [ formula: c₇₀H₁₂₆N₁₂O₁₅(ii) a Accurate quality: 1374.95, respectively; MS (m/z): 1375.63(M +1)⁺、1397.79(M+Na)⁺；TLC R_f: 0.42 (dichloromethane/methanol ═ 9/1); HPLC RT: 12.9min (C8 reverse phase column: 250 mm; acetonitrile/0.077% ammonium acetate in water; operating temperature: 64 ℃ C.; detector: 210nm)]

Example 80

[ (R) - (3- (N, N-dimethylamino) propoxy) methyl-Sar ] -3- [ (gamma-methoxy) -N-MeLeu ] -4-cyclosporin

To [ (R) -alpha-hydroxymethyl-Sar]-3- [ (γ -methoxy) -N-MeLeu]A solution of-4-cyclosporin (250mg, 0.20mmol) in benzene (20ml) was added a solution of sodium hydroxide (633mg, 15.85mmol) in water (0.70ml), tetramethylammonium hydroxide pentahydrate (720mg, 3.96mmol) and 3-dimethylaminopropylchloride hydrochloride (626mg, 3.96 mmol). The mixture was stirred at 40 to 50 ℃ for two days. A solution of sodium hydroxide (633mg, 15.85mmol) in water (0.70ml), tetramethylammonium hydroxide pentahydrate (720mg, 3.96mmol) and 3-dimethylaminopropylchloride hydrochloride (626mg, 3.96mmol) were added and the mixture was kept under stirring for a further two days at 40 to 50 ℃. Another portion of 3-dimethylaminopropyl chloride hydrochloride (1.25g, 7.91mmol) was added and stirring continued for another day at 40 to 50 ℃. A saturated solution of sodium bicarbonate (30ml) was added and the mixture was separated. The aqueous layer was then extracted with ethyl acetate (25 ml. times.2). The combined organic layers were dried over magnesium sulfate and evaporated under reduced pressure. The residue was dissolved in ethyl acetate (25ml), the ethyl acetate phase obtained was washed with acetic acid solution (5ml in 10ml of water) and saturated solution of sodium bicarbonate (30ml), dried over magnesium sulphate and evaporated under reduced pressure. After purification on silica gel, 36mg of product are obtained [ formula: c₆₉H₁₂₆N₁₂O₁₄(ii) a Accurate quality: 1346.95, respectively; MS (m/z): 1347.65(M +1)⁺、1369.74(M+Na)⁺；TLC R_f: 0.21 (dichloromethane/methanol ═ 9/1). HPLC RT: 18.8min (C8 reverse phase column: 250 mm; acetonitrile/0.077% ammonium acetate in water; operating temperature: 64 ℃ C.; detector: 210nm)]。

Example 81

[ (R) - (3- (N, N-diethylamino) propoxy) methyl-Sar ] -3- [ (gamma-methoxy) -N-MeLeu ] -4-cyclosporin

To [ (R) -alpha-hydroxymethyl-Sar]-3- [ (γ -methoxy) -N-MeLeu]A solution of-4-cyclosporin (250mg, 0.20mmol) in 20ml of benzene was added with a solution of sodium hydroxide (633mg, 15.85mmol) in water (0.70ml), followed by tetramethylammonium hydroxide pentahydrate (720mg, 3.96mmol) and 3-diethylaminopropylchloride hydrochloride (737mg, 3.96 mmol). The mixture was stirred at 40 to 50 ℃ for two days. A solution of sodium hydroxide (633mg, 15.85mmol) in water (0.70ml), tetramethylammonium hydroxide pentahydrate (720mg, 3.96mmol) and 3-diethylaminopropyl chloride hydrochloride (737mg, 3.96mmol) were added and the mixture was kept under stirring for a further two days at 40 to 50 ℃. Another portion of 3-diethylaminopropyl chloride hydrochloride (1.47g, 7.91mmol) was added and stirring was continued for a further two days at 40 to 50 ℃. A saturated solution of sodium hydrogencarbonate (30ml) was added and the mixture was separated. The aqueous layer was extracted with ethyl acetate (25 ml. times.2). The combined organic layers were then dried over magnesium sulfate and evaporated under reduced pressure. The residue was dissolved in ethyl acetate (25 ml). The ethyl acetate phase obtained is washed with an acetic acid solution (5ml in 10ml of water) and a saturated solution of sodium bicarbonate (30 ml). Dried over magnesium sulfate and evaporated under reduced pressure. After purification on silica gel, 38mg of product are obtained [ formula: c₇₁H₁₃₀N₁₂O₁₄(ii) a Accurate quality: 1374.98, respectively; MS (m/z): 1375.70(M +1)⁺、1397.80(M+Na)⁺；TLC R_f: 0.24 (dichloromethane/methanol ═ 9/1); HPLC RT: 19.6min (C8 reverse phase column: 250 mm; acetonitrile/0.077% ammonium acetate in water; operating temperature: 64 ℃ C.; detector: 210nm)]

Example 82

[ alpha-methylene-Sar ] -3- [ (gamma-methylthiomethoxy) -N-MeLeu ] -4-Cyclosporin

[ alpha-methylene-Sar ]]-3- [ (γ -methylthiomethoxy) -N-MeLeu]-4-cyclosporin was prepared according to the method described in examples 4 and 5. The product was purified by chromatography on silica gel (ethyl acetate/methanol) [ formula: c₆₅H₁₁₅N₁₁O₁₃S; accurate quality: 1289.84, respectively; MS (m/z): 1290.70(M +1)⁺、1312.67(M+Na)⁺]。

Example 83

[ (R) - (2- (N, N-dimethylamino) ethylthio) methyl-Sar ] -3- [ (gamma-methylthio) methoxy-N-MeLeu ] -4-Cyclosporin

Reacting [ alpha-methylene-Sar]-3- [ (γ -methylthiomethoxy) -N-MeLeu]-4-Cyclosporin (0.32g, 0.25mmol) and 2- (N, N-dimethyl) ethanethiol (0.26g, 2.50mmol) were dissolved in methanol (20ml) and 24 equivalents of triethylamine were added. The mixture was stirred overnight. After removal of the solvent, the residue was subjected to chromatography using dichloromethane/methanol as eluent, yielding 0.14g of pure product [ formula: c₆₉H₁₂₆N₁₂O₁₃S₂(ii) a Accurate quality: 1394.90, respectively; MS (m/z): 1395.70(M +1)⁺、1417.68(M+Na)⁺；TLC R_f: 0.10 (ethyl acetate/methanol-10: 1); HPLC RT: 13.30min (C8 reverse phase column: 250 mm; acetonitrile/water (0.05% trifluoroacetic acid); operating temperature: 64 ℃ C.; detector: 210nm)]。

Example 84

[ (R) - (2- (N, N-diethylamino) ethylthio) methyl-Sar ] -3- [ (gamma-methylthio) methoxy-N-MeLeu ] -4-cyclosporin

Reacting [ alpha-methylene-Sar]-3- [ (γ -methylthiomethoxy) -N-MeLeu]-4-Cyclosporin (0.27g, 0.21mmol) and 2- (N, N-diethyl) ethanethiol (0.28g, 2.10mmol) were dissolved in methanol (20ml), followed by addition of 24 equivalents of triethylamine. The mixture was stirred at room temperature overnight. After removal of the solvent, the residue was purified by chromatography on silica gel using dichloromethane/methanol as eluent to yield 0.17g of pure product [ formula: c₇₁H₁₃₀N₁₂O₁₃S₂(ii) a Accurate quality: 1422.93, respectively; MS (m/z): 1423.70(M +1)⁺、1445.67(M+Na)⁺；TLC R_f: 0.35 (ethyl acetate/methanol-10: 1); HPLC RT: 13.95min (C8 reverse phase column: 250 mm; acetonitrile/water (0.05% trifluoroacetic acid); operating temperature: 64 ℃ C.; detector: 210nm)]。

Example 85

[ Gamma-ethoxymethoxy-N-MeLeu ] -4-cyclosporin

To [ (gamma-hydroxy) -N-MeLeu]A solution of-4-cyclosporin (1.20g, 0.99mmol) in dichloromethane (80ml) was added diisopropylethylamine (FW 129.25, d 0.742, 1.32ml, 0.98g, 7.60mmol), followed by dropwise addition of chloromethyl ethyl ether (FW 94.54, d 1.02, 2.22ml, 2.27g, 24 mmol). The mixture was stirred at room temperature overnight and the completion of the reaction was monitored using TLC. The reaction mixture was washed with 1N hydrochloric acid, saturated aqueous sodium bicarbonate solution and brine. After drying over magnesium sulfate, the mixture was evaporated under reduced pressure to yield a yellowish oil, which was further purified by flash chromatography using dichloromethane/methanol as eluent,0.95g of product [ formula: c₆₅H₁₁₇N₁₁O₁₄(ii) a Accurate quality: 1275.88, respectively; MS (m/z): 1276.70(M + H)⁺、1298.70(M+Na)⁺；TLC R_f: 0.37 (Ethyl acetate)]。

Example 86

[ alpha-methylene-Sar ] -3- [ (gamma-ethoxymethoxy) -N-MeLeu ] -4-Cyclosporin

[ alpha-methylene-Sar ]]-3- [ (γ -ethoxymethoxy) -N-MeLeu]-4-cyclosporin was prepared according to the method described in examples 4 and 5. The product was purified by chromatography on silica gel using ethyl acetate/methanol as eluent [ formula: c₆₆H₁₁₇N₁₁O₁₄(ii) a Accurate quality: 1287.68, respectively; MS (m/z): 1288.72(M +1)⁺、1310.70(M+Na)⁺]。

Example 87

[ (R) - (2- (N, N-diethylamino) ethylthio) methyl-Sar ] -3- [ (gamma-ethoxy) methoxy-N-MeLeu ] -4-cyclosporin

Reacting [ alpha-methylene-Sar]-3- [ (γ -ethoxymethoxy) -N-MeLeu]-4-Cyclosporin (0.27g, 0.21mmol) and 2- (N, N-diethyl) ethanethiol (0.28g, 2.1mmol) were dissolved in methanol (20ml), and 12 equivalents of triethylamine were added. The mixture was stirred at room temperature overnight. After removal of the solvent, the residue was purified by using dichloromethanePurification by chromatography on silica gel using methanol as eluent yielded 90mg of pure product [ formula: c₇₂H₁₃₂N₁₂O₁₄S; accurate quality: 1420.97, respectively; MS (m/z): 1421.75(M +1)⁺、1443.72(M+Na)⁺；TLC：R_f: 0.40 (ethyl acetate/methanol ═ 10/1); HPLC RT: 13.58min (C8 reverse phase column: 250 mm; acetonitrile/water (0.05% trifluoroacetic acid); operating temperature: 64 ℃ C.; detector: 210nm)]。

Example 88

[ alpha-carboxy-Sar ] -3- [ N-MeIle ] -4-Cyclosporin

To a solution of LDA (2.0M in tetrahydrofuran, 5ml, 10mmol) in tetrahydrofuran (15ml) at-78 deg.C under nitrogen was added [ N-MeIle ] over 3min]-a solution of 4-cyclosporin (1.20g, 1.00mmol) 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 allowed to warm slowly to room temperature and kept stirring for another 3 hours. Most of the tetrahydrofuran was evaporated under reduced pressure. Dichloromethane (100ml) and water (50ml) were added. The pH of the mixture was adjusted to around 5 by adding aqueous citric acid. The mixture was then separated and the organic layer was washed with brine, dried over magnesium sulfate and evaporated under reduced pressure to yield 1.10g of crude product which was used in the next step without purification [ formula: c₆₃H₁₁₁N₁₁O₁₄(ii) a Accurate quality: 1245.83, respectively; MS (m/z): 1246.68(M +1)⁺]。

Example 89

[ alpha-methoxycarbonyl-Sar ] -3- [ N-MeIle ] -4-cyclosporin

To [ alpha-carboxy-Sar ]]-3-[N-MeIle]A mixture of-4-cyclosporin (1.00g, 0.80mmol) and potassium carbonate (0.70g, 5.07mmol) in N, N-dimethylformamide (10ml) was added methyl iodide (1.50g, 10.56 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 crude 1.00g of the product [ formula: c₆₄H₁₁₃N₁₁O₁₄(ii) a Accurate quality: 1259.85, respectively; MS (m/z): 1260.51(M +1)⁺]。

Example 90

[ (R) -alpha-hydroxymethyl-Sar ] -3- [ N-MeIle ] -4-cyclosporin

To [ alpha-methoxycarbonyl-Sar ]]-3-[N-MeIle]A suspension of-4-cyclosporin (1.00g, 0.79mmol) and lithium chloride (0.60g, 14.11mmol) in methanol (80ml) was added portionwise to sodium borohydride (3.00g, 79.26 mmol). The mixture was stirred at room temperature overnight. Most of the solvent was evaporated under reduced pressure. Dichloromethane (100ml) 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 97/3) to yield 420mg of product [ formula: c₆₃H₁₁₃N₁₁O₁₃(ii) a Accurate quality: 1231.85, respectively; MS (m/z): 1232.59(M +1)⁺(ii) a TLC Rf: 0.32 (dichloromethane/methanol 95/5); HPLC RT: 14.32min. (C8 reverse phase column: 250 mm; acetonitrile/water(0.05% trifluoroacetic acid); operating temperature: 64 ℃; a detector: 210nm)]。

Example 91

[ alpha-methylene-Sar ] -3- [ N-MeIle ] -4-Cyclosporin

[ alpha-methylene-Sar ]]-3-[N-MeIle]-4-cyclosporin was prepared according to the method described in examples 4 and 5 [ formula: c₆₃H₁₁₁N₁₁O₁₂(ii) a Accurate quality: 1213.84, respectively; MS (m/z): 1214.59(M +1)⁺；TLC R_f: 0.34 (hexane/acetone ═ 6/1); HPLC RT: 17.47min (C8 reverse phase column: 250 mm; acetonitrile/water (0.05% trifluoroacetic acid), operating temperature: 64 ℃ and detector: 210nm]。

Example 92

[ (R) - (2- (N, N-diethylamino) ethylthio) methyl-Sar ] -3- [ N-MeIle ] -4-Cyclosporin

To [ alpha-methylene-Sar ]]-3-[N-MeIle]A solution of-4-cyclosporin (300mg, 0.25mmol) and 2-diethylaminoethanethiol hydrochloride (408mg, 2.41mmol) in methanol (20ml) was added with lithium hydroxide (116mg, 4.83 mmol). The reaction mixture was stirred at room temperature overnight. Most of the solvent was evaporated under reduced pressure. Dichloromethane (80ml) and water (30ml) were added and the mixture was separated. The organic layer was washed with water and brine, dried over magnesium sulfate and evaporated under reduced pressure. The residue was purified by chromatography (dichloromethane/methanol 96 ═ 96/4), 170mg of product are produced [ formula: c₆₉H₁₂₆N₁₂O₁₂S; accurate quality: 1346.93, respectively; MS (m/z): 1347.68(M +1)⁺；TLC R_f: 0.32 (dichloromethane/methanol 95/5); HPLC RT: 13.54min (C8 reverse phase column: 250 mm; acetonitrile/water (0.05% trifluoroacetic acid); operating temperature: 64 ℃ C.; detector: 210nm)]。

Example 93

[ (R) - (2- (N, N-diethylamino) ethoxy) methyl-Sar ] -3- [ N-MeIle ] -4-cyclosporin

To [ (R) -alpha-hydroxymethyl-Sar]-3-[N-MeIle]A solution of-4-cyclosporin (0.39g, 0.32mmol) in benzene (20ml) was added a solution of sodium hydroxide (0.80g, 20mmol) in water (1ml), 2-bromo-N, N-diethylethanamine hydrobromide (2.40g, 9.20mmol) and tetra-N-butylammonium bromide (0.10g, 3.10 mmol). The mixture was stirred at 30 ℃ for 40 hours. Ice water (10ml) was added and the mixture was separated. The aqueous layer was extracted with dichloromethane (50 ml). The combined organic layers were washed with brine, dried over magnesium sulfate and concentrated under reduced pressure. The residue was purified by chromatography (dichloromethane/methanol 96/4) to yield 270mg of product [ formula: c₆₉H₁₂₆N₁₂O₁₃(ii) a Accurate quality: 1330.96, respectively; MS (m/z): 1331.73(M +1)⁺；TLC R_f: 0.34 (dichloromethane/methanol 95/5); HPLC RT: 13.42min (C8 reverse phase column: 250 mm; acetonitrile/water (0.05% trifluoroacetic acid); operating temperature: 64 ℃ C.; detector: 210nm)]。

Example 94

[ alpha-methylene-Sar ] -3- [ N-MeVal ] -4-Cyclosporin

[ alpha ] -methylene group]-3-[N-MeVal]-4-cyclosporin was prepared according to the method described in examples 4 and 5. The product was purified by chromatography on silica gel (ethyl acetate/methanol). [ molecular formula: c₆₂H₁₀₉N₁₁O₁₂(ii) a Accurate quality: 1199.83, respectively; MS (m/z): 1200.56(M +1)⁺、1222.72(M+Na)⁺]。

Example 95

[ (R) - (2- (N, N-dimethylamino) ethylthio) methyl-Sar ] -3- [ N-MeVal ] -4-Cyclosporin

Reacting [ alpha-methylene-Sar]-3-[N-MeVal]-4-Cyclosporin (88mg, 0.07mmol) and 2- (N, N-dimethyl) ethanethiol hydrochloride (0.10g, 7.30mmol) were dissolved in methanol (20ml), followed by addition of 20 equivalents of lithium hydroxide. The mixture was stirred at room temperature overnight. After removal of the solvent, the residue was purified by flash chromatography using dichloromethane/methanol as eluent to yield 30mg of pure product [ formula: c₆₆H₁₂₀N₁₂O₁₂S; accurate quality: 1304.89, respectively; MS (m/z): 1305.68(M +1)⁺、1327.83(M+Na)⁺；TLC R_f: 0.05 (ethyl acetate/methanol ═ 5/1); HPLC RT: 12.23min (C8 reverse phase column: 250 mm; acetonitrile/water (0.05% trifluoroacetic acid); operating temperature: 64 ℃ C.; detector: 210nm)]。

Example 96

[ (R) - (2- (N, N-diethylamino) ethylthio) methyl-Sar ] -3- [ N-MeVal ] -4-cyclosporin

Reacting [ alpha-methylene-Sar]-3-[N-MeVal]-4-Cyclosporin (0.20g, 0.16mmol) and 2- (N, N-diethyl) ethanethiol hydrochloride (0.28g, 1.70mmol) were dissolved in methanol, followed by addition of 20 equivalents of lithium hydroxide. The mixture was stirred overnight. After removal of the solvent, the residue was purified by chromatography on silica gel using dichloromethane/methanol as eluent to yield 100mg of pure product [ formula: c₆₈H₁₂₄N₁₂O₁₂S; accurate quality: 1332.92, respectively; MS (m/e): 1333.58(M +1)⁺、1355.79(M+Na)⁺；TLC R_f: 0.08 (ethyl acetate/methanol ═ 5/1); HPLC RT: 12.77min (C8 reverse phase column: 250 mm; acetonitrile/water (0.05% trifluoroacetic acid); operating temperature: 64 ℃; detector: 210nm)]。

Example 97

[ (R) -alpha-hydroxymethyl-Sar ] -3- [ N-MeVal ] -4-cyclosporin

[ (R) -alpha-hydroxymethyl-Sar]-3-[N-MeVal]-4-cyclosporin was prepared according to the method described in example 2. The product was purified by chromatography on silica gel (ethyl acetate/methanol) [ formula: c₆₂H₁₁₁N₁₁O₁₃(ii) a Accurate quality: 1217.84, respectively; MS (m/z): 1218.56(M +1)⁺、1240.75(M+Na)⁺]。

Example 98

[ (R) - (2- (N, N-dimethylamino) ethoxy) methyl-Sar ] -3- [ N-MeVal ] -4-cyclosporin

To [ (R) -alpha-hydroxymethyl-Sar]-3-[N-MeVal]A solution of-4-cyclosporin (0.12g, 0.10mmol) in benzene (15ml) was added sodium hydroxide (0.20g.5.00mmol), tetramethylammonium hydroxide pentahydrate (0.18g, 1.00mmol) and 3-dimethylaminoethyl chloride hydrochloride (0.14g, 1.00 mmol). The mixture was stirred at 30 ℃ overnight. Ice water (20ml) was added and the mixture was separated. The aqueous layer was extracted with ethyl acetate (20 ml). The combined organic layers were washed with brine, dried over magnesium sulfate and evaporated under reduced pressure. The residue was subjected to chromatography on silica gel (dichloromethane/methanol ═ 95/5) yielding 30mg of pure product [ formula: c₆₆H₁₂₀N₁₂O₁₃(ii) a Accurate quality: 1288.91, respectively; MS (m/z): 1289.73(M +1)⁺、1311.71(M+Na)⁺；TLC R_f: 0.14 (dichloromethane/methanol ═ 10/1); HPLC RT: 12.00min (C8 reverse phase column: 250 mm; acetonitrile/water (0.05% trifluoroacetic acid); operating temperature: 64 ℃; detector: 210nm)]。

Example 99

[ (R) -alpha- (N-piperidinyl) methyl-Sar ] -3- [ (gamma-hydroxy) -NMeLeu ] -4-cyclosporin

In the presence of catalytic amount of copper (II) acetate [ alpha-methylene-Sar ]]-3- [ (γ -hydroxy) -NMeLeu]-4-Cyclosporin (0.37g, 0.30mmol) andpiperidine (0.26g, 3.00mmol) was dissolved in acetonitrile/water (20 ml). The mixture was stirred at room temperature overnight. After removal of the solvent, the residue was dissolved in dichloromethane (30 ml). The dichloromethane phase was washed with brine, dried over magnesium sulfate and evaporated under reduced pressure. The residue was further purified by chromatography on silica gel (dichloromethane/methanol, 96/4) to yield 0.17g of product [ formula: c₆₈H₁₂₂N₁₂O₁₃(ii) a Accurate quality: 1314.93, respectively; MS (m/z): 1315.74(M +1)⁺、1337.86(M+Na)⁺；TLC R_f: 0.10 (ethyl acetate/methanol ═ 5/1); HPLC RT: 11.70min (C8 reverse phase column: 150 mm; acetonitrile/water (0.05% TFA); operating temperature: 64 ℃; detector: 210nm)]。

Reference example 1

[ alpha-carboxy-Sar ] -3-cyclosporin

N-butyllithium (2.87M, 27mmol, 9.4ml, 10eq) was added to a solution of diisopropylamine (3.8ml, 27mmol, 10eq) in tetrahydrofuran (80ml) at-78 ℃ under nitrogen. After stirring the reaction mixture for one hour, a solution of cyclosporin A (3.2g, 2.66mmol) in tetrahydrofuran (15ml) was added over 10min. Stirring was continued for 2 hours at-78 ℃. Carbon dioxide gas was bubbled through the reaction mixture for 20-25 minutes and stirred at-78 ℃ for another hour. The cooling bath was then removed and the reaction mixture was allowed to slowly warm to 0 ℃. Most of the tetrahydrofuran was removed under vacuum at room temperature. The residue was quenched by addition of saturated citric acid solution and the pH of the mixture was adjusted to around 7-8. Unreacted cyclosporine was extracted with ether (40 ml. times.2). 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 a semi-solid product (2.61g, yield: 78%) [ formula: c₆₃H₁₁₁N₁₁O₁₄(ii) a Accurate quality: 1245.83, respectively; MS (m/z): 1246.7(M +1)⁺、1268.7(M+Na)⁺]。

[ α -carboxy-Sar ] -3-cyclosporin was synthesized according to the procedure described by Seebach D, et al, 1993, Helv ChimActa, 76, 1564-1590.

Reference example 2

Synthesis of [ N-MeVal ] -4-Cyclosporin (SDZ-220-384)

[ N-MeVal ] -4-Cyclosporine (SDZ 220-384) was synthesized according to the procedure described by Papageorgiou C, et al, 1994, Bioorg & Med Chem Lett, 4, 267-272 and its key cyclosporine ring opening between positions 3 and 4 was as cited in reference 14: su Z and Wenger R, unpublished results; papageorgiou C, et al, 1994, j.med.chem., 37, 3674-3676 and its key cyclosporine opening between positions 3 and 4 as cited in reference 11: su Z and Wenger R, unpublished results.

Cyclosporin A-acetate salt

To cyclosporin A: (1) (12.00g, 19.98mmol) in acetic anhydride (MW: 102.09, d1.082, 40ml) was added pyridine (MW: 79.01, d 0.978, 40ml) and 4-N, N-dimethylaminopyridine (MW: 122.17, 0.40 g). The mixture was stirred at room temperature overnight, and then the mixture was diluted with 600ml of ethyl acetate. The mixture was washed with brine, saturated ammonium chloride solution and 15% sodium bicarbonate solution. The organic phase is dried over sodium sulfate, filtered and evaporated under reduced pressure. All pyridine was then removed by azeotropic evaporation under reduced pressure by adding toluene to the mixture, yielding a pale yellow solid residue which was purified by flash chromatography on a silica gel column (100-200 mesh) using an eluent of ethyl acetate/hexane (1/3)This gave 11.80g (9.48mmol, 95%) of cyclosporin A-acetate (2)。

MeLeuValMeLeuAlaDAlaMeLeuMeLeuMeValMeBmt(OAc)AbuSar-Ome

To a suspension of trimethyloxonium-fluoroboric acid (MW: 147.91, 2.96g, 20mmol, 2.50 equiv.) in dichloromethane (80m1) was added cyclosporin A-acetate (2) (10.00g, 8.00 mmol). The suspension was stirred at room temperature for 18 h, then a solution of sodium methoxide (9.90mmol) in methanol (40ml) was added. After stirring the mixture for another half hour, a 2N solution of sulfuric acid in methanol (40ml) was added. The mixture was stirred at room temperature for 15-30 minutes and neutralized with 15% potassium bicarbonate solution. The mixture is then extracted twice with 700ml of ethyl acetate. The combined organic layers were washed with brine, dried over sodium sulfate and evaporated under reduced pressure. The residue was purified by flash chromatography on a silica gel column (100-200 mesh) using an eluent of methanol/methyl tert-butyl ether to give 7.15g (5.60mmol, 70%) of linear undecapeptide (R) ((R))3)。

phenylthiourea-MeLeuValMeLeuAlaDALaMeLeuMeLeuMeLeuMeValMeBmt (OAc) AbuSar-Ome

To a linear undecapeptide (3) (7.00g, 5.50mmol) in tetrahydrofuran (80ml) was added phenyl isothiocyanate (MW: 135.19, d 1.130, 0.86ml, 7.15mmol, 1.30 equiv). The mixture was stirred at room temperature for 3 hours and evaporated under reduced pressure. The residue was purified by flash chromatography on a silica gel column (100-200 mesh) using an eluent of acetone/hexane (1/5) to give 6.99g (4.95mmol, 90%) of linear phenylthiourea undecapeptide (4) [ accurate quality: 1410.89, respectively; MS m/z: 1433.88 (M)+Na)⁺]。

ValMeLeuAlaDAlaMeLeuMeLeuMeValMeBmt(OAc)AbuSar-Ome

To linear phenylthiourea undecapeptides (f) at room temperature4) (6.80g, 4.82mmol) in toluene (300ml) was added trifluoroacetic acid (MW: 114.02, d 1.480, 8.00 ml). The mixture was stirred for 1.5 to 2 hours and quenched by a slurry of sodium bicarbonate in water. The mixture was then separated and the aqueous phase was extracted with toluene (100ml) and then ethyl acetate (100 ml). The combined organic layers were dried over sodium sulfate and evaporated under reduced pressure. The residue was purified by flash chromatography on a silica gel column (100-200 mesh) using an eluent of acetone/hexane (3/1) to give 3.88g (3.37mmol, 70%) of a linear decapeptide ((3.37 mmol, 70%))5) [ accurate quality: 1148.78, respectively; MS m/z: 1149.78(M +1)⁺]。

This Edman degradation was performed according to a similar method described by Edman P, et al, 1967, eur.j. biochem., 1, 80.

BocMeValValMeLeuAlaDAlaMeLeuMeLeuMeValMeBmt(OAc)AbuSar-Ome

Linear decapeptide at 0 ℃ (C)5) (3.80g, 3.30mmol) in dichloromethane (150ml) Boc-MeVal (M-methyl-tert-butyl-N-methyl-ethyl-N-methyl-tert-butyl-N-methyl-N-6) (MW: 231.29, 0.92g, 3.96mmol, 1.2 eq), 1-propanephosphonic acid cyclic anhydride (MW: 318.18, 2.10ml, 50 wt.% solution in ethyl acetate) and triethylamine (MW: 101.19, d 0.726, 0.46ml, 3.30 mmol). The resulting mixture was stirred at room temperature for 5 hours. The mixture was then washed with brine. The aqueous layer was extracted with ethyl acetate (100 ml). The combined organic layers were dried over sodium sulfate. Removing the solvent under reduced pressureThe reagent yielded a residue that was purified by flash chromatography on a silica gel column (100-200 mesh) using an eluent of acetone/hexane (1/2.5) yielding 4.05g (2.97mmol, 90%) of a linear Boc-N-MeVal-decapeptide (Sigma) ((2.97 mmol, 90%))7) [ accurate quality: 1361.91, respectively; MS m/z: 1384.91(M + Na)⁺]。

BocMeValValMeLeuAlaDAlaMeLeuMeLeuMeValMeBmt(OAc)AbuSar-OH

Linear [ Boc-N-MeVal at 0 ℃]-4-decapeptide (7) (4.00g, 2.94mmol) in ethanol (150ml) was added 0.5N sodium hydroxide solution (7.1ml, 1.20 equiv). The mixture was stirred and held at 0 ℃ for 16 hours, then the pH of the mixture was adjusted to around 3 by adding 0.5N hydrochloric acid. Most of the solvent was evaporated under reduced pressure and the residue was dissolved in 200ml of ethyl acetate. The mixture was washed with pH3 buffer, dried over sodium sulfate, filtered and evaporated under reduced pressure. The residue was purified by flash chromatography on a silica gel column (100-200 mesh) using an eluent of methanol/ethyl acetate (1/8) to give 2.55g (1.89mmol, 64.3%) of the free acid (R) ()8)。

MeValValMeLeuAlaDAlaMeLeuMeLeuMeValMeBmt(OAc)AbuSar-OH

To the free acid at 0 ℃: (8) (2.55g, 1.89mmol) in dichloromethane (25ml) 5ml of trifluoroacetic acid (MW: 114.02, d 1.480). The solution was stirred at room temperature for 2 hours. Ethyl acetate (300ml) was then added and the solvent was removed under reduced pressure. Additional ethyl acetate (300ml) was added and the solvent was again removed under reduced pressure. The residue was purified by flash chromatography on a silica gel column (100-200 mesh) using an eluent of methanol/acetone (1/3) to yield 2.01g (1.61mmol, 85%) of Linear [ N-MeVal]-4-decapeptide free acid (9) [ accurate quality: 1247.85, respectively; MS m/z: 1248.85(M +1)⁺]。

[ N-MeVal ] -4-Cyclosporine acetate

Linear [ N-MeVal ] at 0 deg.C]-4-decapeptide free acid (9) (1.03g, 0.83mmol) in dichloromethane (250ml) 1-propanephosphonic acid cyclic anhydride (MW: 318.18, 0.53ml, 50 wt.% solution in ethyl acetate), 2, 4, 6-trimethylpyridine (MW: 121.18, d 0.917, 0.11ml, 0.83 mmol)). The mixture was stirred at room temperature for 24 hours. The mixture was then passed through a thin layer of silica gel and washed twice with 40ml of ethyl acetate. The collected organic solution was evaporated under reduced pressure. The residue was purified by flash chromatography on a silica gel column (230-400 mesh) using an eluent of methanol/acetone (1/6) to give 611mg (0.50mmol, 60%) of (N-methyl-Val) -4-cyclosporine acetate (R) ((0.50 mmol, 60%))10) [ accurate quality: 1229.84, respectively; MS m/z: 1252.82(M + Na)⁺]。

[ N-MeVal ] -4-Cyclosporin

To [ N-MeVal]-4-Cyclosporine acetate salt (10) (0.60g, 0.49mmol) in methanol (40ml) was added a solution of sodium methoxide in methanol (0.5M, 1.9ml, 2.0 equiv). The mixture was stirred at 0 ℃ for 0.5 h and at room temperature for 24 h. The pH of the mixture was adjusted to around 6 by adding 0.5N hydrochloric acid. After evaporation of the solvent under reduced pressure, the residue was dissolved in 200ml of ethyl acetate. The organic solution was washed with aqueous sodium bicarbonate and brine, dried over sodium sulfate and filtered. After removal of the solvent, the residue was purified by using acetone/hexane (1/2)) Purification by flash chromatography on a silica gel column (230-400 mesh) to yield 406mg (0.34mmol, 70%) of [ N-MeVal ]]-4-Cyclosporin (c)11) [ accurate quality: 1187.83, respectively; 84; MS m/z: 1210.81(M + Na)]。

Reference example 3

[ N-MeIle ] -4-cyclosporin (NIM-811) was prepared according to the procedure used for the synthesis of (N-MeVal) -4-cyclosporin (SDZ 220-384).

Reference example 4

Side chain intermediates were synthesized according to the procedure described by Urquhart GG, 1994, Org Synth, col. vol III, 36

2-morpholinylethylthiol

A mixture of 4- (2-chloroethyl) morpholine (7.00g, 37mmol) and thiourea (2.90g, 38mmol) in 95% ethanol (55ml) was heated at reflux for 24 h. A solution of sodium hydroxide (3.40g, 85mmol) in water (20ml) was added and the mixture was continued to reflux for a further 3 hours. After cooling to room temperature, the mixture was evaporated under reduced pressure. The residue was mixed with benzene. The benzene layer was washed with brine, dried over magnesium sulfate and evaporated to give 3.80g of crude product, which was used for the addition reaction.

2- (N-piperidinyl) ethanethiol

A mixture of 1- (2-chloroethyl) piperidine hydrochloride (7.00g, 38mmol) and thiourea (4.60g, 61mmol) in 95% ethanol (30ml) was heated under reflux for 24 h. A solution of sodium hydroxide (2.40g) in water (20ml) was added. The mixture was continued to reflux for another 3 hours. After cooling to room temperature, the mixture was evaporated under reduced pressure. The residue was mixed with ether. The organic layer was washed with brine, dried over magnesium sulfate and evaporated under reduced pressure to give 3.20g of crude product, which was used for addition reaction without purification.

2- (N-pyrrolidinyl) ethanethiol

A mixture of 1- (2-chloroethyl) piperidine hydrochloride (7.0g, 41mmol) and thiourea (3.20g, 40mmol) in 95% ethanol (30ml) was heated under reflux for 24 h. A solution of sodium hydroxide (3.40g, 85mmol) in water (20ml) was added. The mixture was continued to reflux for another 3 hours. After cooling to room temperature, the mixture was evaporated under reduced pressure. The residue was mixed with benzene. The organic layer was washed with brine, dried over magnesium sulfate and evaporated under reduced pressure to give 3.80g of crude product, which was used for addition reaction without purification.

3- (N-pyrrolidinyl) propanethiol

To a suspension of 1-bromo-3-chloropropane (30.00g, 191mmol) and potassium carbonate (17.00g, 123mmol) in dichloromethane (160ml) was added pyrrolidine (3.50g, 49mmol) in portions. The mixture was stirred at room temperature overnight. The mixture was then filtered and evaporated under reduced pressure. The residue was purified by chromatography (ethyl acetate/methanol 95/5) to yield 6.00g of product.

A mixture of 1- (3-chloropropyl) pyrrolidine (3.4g, 23mmol) and thiourea (1.8g, 23mmol) in 95% ethanol (55ml) was heated at reflux for 24 h. A solution of sodium hydroxide (1.20g, 30mmol) in water (10ml) was added and the mixture was continued to reflux for a further 3 hours. After cooling to room temperature, the mixture was evaporated under reduced pressure. The residue was mixed with benzene. The organic layer was washed with brine, dried over magnesium sulfate and evaporated under reduced pressure to give 1.80g of crude product, which was used for the addition reaction.

3- (N-piperidinyl) propanethiol

A mixture of 1- (3-chloropropyl) piperidine hydrochloride (7.50g, 38mmol) and thiourea (4.60g, 61mmol) in 95% ethanol (30ml) was heated at reflux for 24 h. A solution of sodium hydroxide (2.40g) in water (20ml) was added. The mixture was continued to reflux for another 3 hours. After cooling to room temperature, the mixture was evaporated under reduced pressure. The residue was mixed with ether. The organic layer was washed with brine, dried over magnesium sulfate and evaporated under reduced pressure to give 3.50g of crude product, which was used for addition reaction without purification.

3-morpholinyl propanethiol

To a suspension of 1-bromo-3-chloropropane (30.00g, 191mmol) and potassium carbonate (14.00g, 101mmol) in dichloromethane (160ml) was added morpholine (4.00g, 46mmol) in portions. The mixture was then stirred at room temperature overnight. The mixture was filtered and evaporated under reduced pressure. The residue was purified by chromatography (ethyl acetate) to yield 5.60g of product.

A mixture of 1- (3-chloropropyl) morpholine (4.20g, 25.76mmol) and thiourea (2.00g, 26.27mmol) in 95% ethanol (55ml) was heated at reflux for 24 h. A solution of sodium hydroxide (1.3g, 32.50mmol) in water (10ml) was added and the mixture was continued to reflux for a further 3 hours. After cooling to room temperature, the mixture was evaporated under reduced pressure. The residue was mixed with benzene. The benzene layer was washed with brine, dried over magnesium sulfate and evaporated under reduced pressure to give 2.20g of crude product, which was used for the addition reaction.

2- (4-methyl-N-piperazinyl) ethanethiol

A mixture of 2- (4-methylpiperazine) ethyl chloride (8.00g, 40mmol) and thiourea (4.87g, 64mmol) in 95% ethanol (55ml) was heated under reflux for 24 hours. A solution of sodium hydroxide (2.60g) in water (20ml) was added and the mixture was continued to reflux for a further 3 hours. After cooling to room temperature, the mixture was evaporated under reduced pressure. The residue was mixed with benzene. The benzene layer was washed with brine, dried over magnesium sulfate and evaporated to give 3.0g of crude product, which was used for the addition reaction.

3- (4-methyl-N-piperazinyl) propanethiol

A mixture of 3- (4-methylpiperazine) propyl chloride (8.5g, 40mmol) and thiourea (4.87g, 64mmol) in 95% ethanol (70ml) was heated under reflux for 24 hours. A solution of sodium hydroxide (2.6g) in water (20ml) was added and the mixture was continued to reflux for a further 3 hours. After cooling to room temperature, the mixture was evaporated under reduced pressure. The residue was mixed with ether. The organic layer was washed with brine, dried over magnesium sulfate and evaporated to give 2.5g of crude product, which was used for the addition reaction.

3- (N-Ethyl-N-isopropylamino) propanethiol

To a suspension of 1-bromo-3-chloropropane (11.00g, 70mmol) and potassium carbonate (13.00g, 94mmol) in dichloromethane (100ml) was added ethyl isopropylamine (4.10g, 47mmol) in portions. The mixture was stirred at room temperature overnight. The mixture was filtered and concentrated under reduced pressure. The residue was purified by chromatography (ethyl acetate/methanol 95/5) to yield 6.10g of product.

A mixture of 3-chloropropylethylisopropylamine (4.20g, 25.66mmol) and thiourea (2.00g, 26.32mmol) in 95% ethanol (55ml) was heated under reflux for 24 h. A solution of sodium hydroxide (1.30g, 32.50mmol) in water (10ml) was added and the mixture was continued to reflux for a further 3 hours. After cooling to room temperature, the mixture was evaporated under reduced pressure. The residue was mixed with benzene. The benzene layer was washed with brine, dried over magnesium sulfate and evaporated under reduced pressure to give 1.20g of crude product, which was used for the addition reaction.

Example 100

Stability tests of [ (R) -3- (N, N-dimethylamino) ethylthio-Sar ] -3- [ (γ -hydroxy) -NMeLeu ] -4-Cyclosporin (SCY-635) and Cyclosporin derivatives

Cyclosporine derivatives were evaluated for stability in methanol at 65 ℃ and 50 ℃ and HPLC was used to monitor possible isomerisation of these compounds. SCY-635 was found to be unstable and readily converted to its corresponding epimer, which is expected to have low or no antiviral activity.

SCY-635^*Epimerization in MeOH at 65 deg.C

^*SCY-635 was prepared according to the method described below: evans M, et al,2003, bioorg.med.chem.lett., 4, 4415-4419; carry J, et al, 2004, Synlett.2, 316-320; or U.S. patent No. 5,994,299 (each of which is incorporated herein by reference).

Epimer of SCY-635^*Epimerization in MeOH at 65 deg.C

^*During the stability study, it was found that SCY-635 was converted to its epimer, which was isolated as a pure compound. HPLC RT: 14.60 minutes (SCY-635) and: 15.01 min (epimer thereof) (C8 reverse phase column, 250mm, acetonitrile/0.077% NH)₄OAc in water, operating temperature: 64 ℃; a detector: 210 nm).

When this epimer was treated with MeOH at 65 deg.C, it was also found to be partially converted to SCY-635. At the end of the equilibrium, the solution contained about 58% SCY-635 and about 42% epimer.

TABLE 1 epimerization of [ (R) -2- (N, N-dimethylamino) ethylthio-Sar ] -3-cyclosporin in MeOH at 65 deg.C

TABLE 2 epimerization of [ (R) -3- (N-morpholinyl) propylthio-Sar ] -3- [ (γ -hydroxy) -NMeLeu ] -4-Cyclosporin in MeOH at 65 deg.C

TABLE 3 epimerization of [ (R) -3- (N-morpholinyl) propylthio-Sar ] -3- [ (γ -methoxy) -NMeLeu ] -4-Cyclosporin in MeOH at 65 deg.C

TABLE 4 epimerization of cyclosporin derivatives in MeOH at 50 deg.C

TABLE 5 epimerization of cyclosporin derivatives in MeOH at 50 deg.C-58 deg.C

Based on the isomerization data, the inventors propose that epimerization of SCY-635 occurs by the following process:

thus, the two carbon side chains at position 3 of the sarcosine of cyclosporine promote instability as it can form a six-membered ring transition state and stimulate epimerization. In addition, epimerization is accelerated by the gamma-hydroxy group at the 4-position of leucine.

Thus, the inventors have conceived novel cyclosporin derivatives having enhanced stability while maintaining good cyclophilin binding activity. In particular, the inventors have surprisingly found that masking the γ -hydroxy group on the leucine at position 4, extending the side carbon chain (e.g., with 3 carbons or higher), and/or substituting the amine terminus at position 3 with a bulky side chain can prevent or reduce epimerization. In particular, when a methylene substituent is introduced at position-3, those analogs are very stable and can prevent epimerization.

Example 101

anti-HCV activity of cyclosporin derivatives

Cyclosporine derivatives were evaluated for anti-HCV activity in an HCV subgenomic replicon assay. The assay used the cell line ET (Luc-ubi-neo/ET), which is the Huh7 human hepatoma cell line carrying HCV replicons with stable luciferase (Luc) reporters. HCV RNA replication was assessed by quantifying HCV replicon-derived luciferase activity. Derivation of EC by assessment of antiviral Activity of Cyclosporin analogs Using luciferase Ends after drug treatment₅₀And EC₉₀Values Krieger, n., et al, 2001, j.virol.75, 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 6 test results for certain representative compounds

TABLE 7 test results for certain representative compounds

TABLE 8 test results for certain representative compounds

TABLE 9 test results for certain representative compounds

Example 102

anti-HIV activity of cyclosporin derivatives

In the use of CEM-SS cells and HIV-1_IIIBOr HIV-1RF, by cytoprotective assays. Resist againstViral activity is measured as a reduction in the cytopathic effect caused by the virus when the compound prevents viral replication. Cytoprotection and compound cytotoxicity was assessed using the tetrazolium dye MTS (Promega) to calculate cell viability after 6-day incubation following viral infection (Zhou g., et al, 2011, j.med.chem.27, 7220-31; incorporated herein by reference).

TABLE 10 test results (MTS endpoint) of certain representative compounds against HIV-1IIIB in CEM-SS cells

Examples 103 to 705

Antiviral cyclosporin derivatives

The following compounds may be prepared according to methods analogous to those described herein.

TABLE 11

TABLE 12

Watch 13

TABLE 14

Watch 15

Claims

1. A compound of formula (I):

or a pharmaceutically acceptable salt thereof, wherein:

R₈is n-butyl, (E) -but-2-enyl or

R₂Is ethyl, 1-hydroxyethyl, isopropyl or n-propyl;

w is O, S or NR₁；

R₁Is hydrogen;

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

(C₃-C₇) Cycloalkyl, optionally with (C)₁-C₆) Alkyl substitution;

R₃the method comprises the following steps:

H；

R₇is that

R₅The method comprises the following steps:

H；

each occurrence of R_AAnd R_BIndependently are:

hydrogen;

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

(C₃-C₇) Cycloalkyl, optionally with (C)₁-C₆) Alkyl substitution;

each occurrence of R_CIndependently is hydrogen or (C)₁-C₆) An alkyl group;

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

m is an integer of 1, 2, 3, 4 or 5.

2. The compound of claim 1, wherein R₈Is n-butyl.

3. The compound of claim 1, wherein R₈Is (E) -but-2-enyl.

4. The compound of any one of claims 1-3, wherein R₂Is ethyl.

5. The compound of claim 1, having the structure of formulae (II) to (V):

or a pharmaceutically acceptable salt thereof, wherein:

represents a single or double bond;

each W is independently O, S or NR₁；

Each R₁Independently is hydrogen;

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

(C₃-C₇) CycloalkanesOptionally with (C)₁-C₆) Alkyl substitution;

each R₃Independently are:

H；

(C₂-C₆) Alkenyl, optionally substituted by one or more substituents which may be the same or different, 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_AToOne or more groups;

each R₅Independently are:

H；

each occurrence of R_AAnd R_BIndependently are:

hydrogen;

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

(C₃-C₇) Cycloalkyl, optionally with (C)₁-C₆) Alkyl substitution;

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’；

Each occurrence of R_CIndependently is hydrogen or (C)₁-C₆) An alkyl group;

each occurrence of R_FAnd R_F’Independently of each other, 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 three to seven ring atoms, which ring may optionally contain another heteroatom selected from the group consisting of nitrogen, oxygen and sulfur and may optionally be substituted by one to four groups selected from the group consisting of alkyl, phenyl and benzyl, which may be the same or different;

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

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

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

6. The compound of any one of claims 1-5, wherein W is O.

7. The compound of any one of claims 1-5, wherein W is S.

8. The compound of any one of claims 1-5, wherein W is NR₁。

9. The compound of any one of claims 1-5, wherein W is NH.

10. The compound of any one of claims 1-5, wherein W is N (C)₁-C₄) An alkyl group.

11. The compound of any one of claims 1-10, wherein R₃Is H, methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, tert-butyl, CH₂CMe₃Phenyl, CH₂-phenyl group,

12. The compound of any one of claims 1-10, wherein R₃Is- (CH)₂)_nNR_AR_BWherein n is an integer of 1, 2, 3, 4, 5 or 6; and wherein R is at each occurrence_AAnd R_BIndependently is hydrogen; (C)₁-C₄) Alkyl, optionally substituted by one or more radicals R which may be the same or different_DSubstituted, wherein R is at each occurrence_DIndependently 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 three to seven ring atoms, which ring optionally contains another heteroatom selected from the group consisting of nitrogen, oxygen and sulfur and may optionally be selected from (C), which may be the same or different₁-C₄) Alkyl, phenyl and benzyl.

13. The compound of any one of claims 1-10, wherein R₃Is- (CH)₂)_nNR_AR_BWherein n is an integer of 1, 2, 3, 4, 5 or 6; and wherein R_AAnd R_BTogether with the nitrogen atom to which they are attached, form a saturated or unsaturated heterocyclic ring containing three to seven ring atoms, which optionally contains another heteroatom selected from nitrogen, oxygen and sulfur and which optionally may be selected from (C), which may be the same or different₁-C₄) Alkyl, phenyl and benzyl.

14. The compound of any one of claims 1-10, wherein R₃Is 2-aminoethyl, 2-aminobutyl, 3-aminobutyl, 2-monoalkylaminoethyl, 2-monoalkylaminobutyl, 3-monoalkylaminobutyl, 2-dialkylaminoethyl, 2-dialkylaminobutyl or 3-dialkylaminobutyl, wherein the alkyl is (C)₁-C₄) An alkyl group.

15. The compound of any one of claims 1-10, wherein R₃Is dimethylaminoethyl, diethylaminoethyl, methylethylaminoethyl, methyl-iso-butylaminoethyl, ethyl-iso-butylaminoethyl, methyl-tert-butylaminoethyl or ethyl-tert-butylaminoethyl.

16. The compound of any one of claims 1-10, wherein R₃The method comprises the following steps:

wherein n is an integer of 2, 3, 4, 5 or 6 and m is an integer of 2, 3 or 4.

17. The compound of any one of claims 1-16, wherein 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 one to three substituents of halogen; 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 of 0, 1, 2, 3, 4 or 5; and m is an integer of 1, 2, 3, 4 or 5.

18. The compound of any one of claims 1-17, wherein R at each occurrence_AAnd R_BIndependently 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.

19. The compound of any one of claims 1-17, wherein 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.

20. The compound of any one of claims 5-19, whereinRepresents a single bond.

21. The compound of any one of claims 5-19, whereinRepresents a double bond.

22. The compound of claim 1, having the structure of formulae (IIa) - (Va):

or a pharmaceutically acceptable salt thereof, wherein:

represents a single or double bond;

each W is independently O, S or NR₁；

Each R₁Independently is hydrogen;

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

(C₃-C₇) Cycloalkyl, optionally with (C)₁-C₆) Alkyl substitution;

each R₅Independently are:

H；

each occurrence of R_AAnd R_BIndependently are:

hydrogen;

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

(C₃-C₇) Cycloalkyl, optionally with (C)₁-C₆) Alkyl substitution;

or R_AAnd R_BTogether with the nitrogen atom to which they are attached, form a saturated or unsaturated heterocyclic ring containing three to seven ring atoms, which optionally contains another heteroatom selected from the group consisting of nitrogen, oxygen and sulfur and which optionally may be the same or differentOne to four groups selected from the group consisting of alkyl, phenyl and benzyl;

each occurrence of R_CIndependently is hydrogen or (C)₁-C₆) An alkyl group;

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

Each of m, n, and q is independently an integer of 1, 2, 3, 4, or 5.

23. The compound of claim 1, having the structure of formulae (IIb) - (Vb):

or a pharmaceutically acceptable salt thereof, wherein:

represents a single or double bond;

each R₁Independently is hydrogen;

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

(C₃-C₇) Cycloalkyl, optionally with (C)_1-C₆) Alkyl substitution;

each R₃Independently are:

H；

(C₃-C₇) Cycloalkyl, optionally substituted by a substituent which may be the same or different, selected from halogen, hydroxy,one or more groups of amino, monoalkylamino and dialkylamino;

each R₅Independently are:

H；

each occurrence of R₄Independently halogen, hydroxy, aryl (e.g. phenyl), O (CH)₂)_mOR_A、O(CH₂)_mO(CH₂)_mOR_A、C(＝O)(C₁-C₆) Alkyl, C (═ O) OR_A、C(＝O)NR_AR_B、-NR_AR_B、-MR_CCH₂(CH₂)_pNR_AR_B、NR_CCH₂(CH₂)_pNR_A]_mCH₂(CH₂)_nNR_AR_B、O[CH₂(CH₂)_pO]_mCH₂(CH₂)_nOR_A、OCH₂(CH₂)_pNR_AR_BOr O [ CH ]₂(CH₂)_pO]_mCH₂(CH₂)_nNR_AR_B；

each occurrence of R_AAnd R_BIndependently are:

hydrogen;

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

(C₃-C₇) Cycloalkyl, optionally with (C)₁-C₆) Alkyl substitution;

each occurrence of R_CIndependently is hydrogen or (C)₁-C₆) An alkyl group;

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

Each of m, n, and q is independently an integer of 1, 2, 3, 4, or 5.

24. The compound of claim 23, wherein R₁Is hydrogen or (C)₁-C₆) Alkyl, and R₃Is (C)₁-C₆) An alkyl group.

25. The compound of claim 23, wherein R₁And R₃Together 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.

26. The compound of claim 5, wherein:

represents a single or double bond;

each W is independently O, S or NR₁；

Each R₁Independently is hydrogen;

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

(C₃-C₇) Cycloalkyl, optionally with (C)₁-C₆) Alkyl substitution;

phenyl, optionally substituted with one or more substituents which may be the same or different, selected from halogen, -O (C)₁-C₆) Alkyl, -C (═ O) O (C)₁-C₆) Alkyl, amino, alkylamino and dialkylaminoOne to five groups of radicals;

each R₃Independently is

R₅The method comprises the following steps:

H；

(C₃-C₇) Cycloalkyl, optionally substituted by one or more substituents which may be the same or differentDifferent substitution of one or more groups selected from halogen, hydroxy, amino, monoalkylamino and dialkylamino;

each occurrence of R_AAnd R_BIndependently are:

hydrogen;

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

(C₃-C₇) Cycloalkyl, optionally with (C)₁-C₆) Alkyl substitution;

each occurrence of R_CIndependently is hydrogen or (C)₁-C₆) An alkyl group;

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

Each of m, n, and q is independently an integer of 1, 2, 3, 4, or 5.

27. The compound of claim 22, wherein q is 1 or 2.

28. The compound of any one of claims 22-27, wherein W is O.

29. The compound of any one of claims 22-27, wherein W is S.

30. The compound of any one of claims 22-27, wherein W is NH.

31. The compound of any one of claims 22-27, wherein W is N- (C)₁-C₄) An alkyl group.

32. The compound of any one of claims 22-31, wherein:

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 one to three substituents of halogen; 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 of 0, 1, 2, 3, 4 or 5; and m is an integer of 1, 2, 3, 4 or 5.

33. The compound of any one of claims 22-31, wherein:

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.

34. The compound of any one of claims 22-31, wherein R₅Is H.

35. The compound of any one of claims 22-31, wherein R₅Is methyl.

36. The compound of any one of claims 22-35, wherein R at each occurrence_AAnd R_BIndependently 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.

37. According to the rightThe compound of any one of claims 22-35, wherein R at each occurrence_AAnd R_BIndependently is H or (C)₁-C₆) An alkyl group.

38. The compound of any one of claims 22-35, wherein 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.

39. A compound selected from:

40. A compound having the structure:

or a pharmaceutically acceptable salt thereof.

41. A pharmaceutical composition comprising at least one compound according to any one of claims 1-36 and a pharmaceutically acceptable carrier or diluent.

42. 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 of any one of claims 1-36.

43. 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 of any one of claims 1-36.

44. A method of 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 1-36.