TW200302105A - Use of PDE5 inhibitors in the treatment of scarring - Google Patents

Abstract

This invention relates to the use of selective cyclic guanosine 3', 5'-monophosphate type five (cGMP PDE5) inhibitors (hereinafter PDE5 inhibitors), including in particular the compound sildenafil, for the treatment of or prevention of scarring or fibrosis in tissue.

Description

200302105 (1) 发明 Description of the invention [Technical field to which the invention belongs] The present invention relates to cyclic guanine nucleoside 3,5 · -phosphate fifth type cGMP PDE5 inhibitor (hereinafter referred to as PDE 5 inhibitor), including , Especially the compound sildenafil, for use in reducing or preventing scar formation and / or fibrosis. Examples of diseases related to scar formation and / or fibrosis according to the present invention include (but are not necessarily limited to): pulmonary fibrosis, atherosclerosis, cardiovascular disease, infections, trauma, surgery, or thermal injury. Scar formation and / or fibrosis of the skin and cornea, scleroderma and other connective tissue disorders, fibrosis of the heart, chronic obstructive pulmonary disease, muscle fibrosis, renal fibrosis, chronic skin ulcers and lipdermatosclerosis , Pulmonary fibrosis or any origin), post-operative and spontaneous adhesions' skin inflammation (including moss and related conditions), aging and all aging-related degeneration disorders (including skin aging), liver fibrosis or Any cause (including viral and non-viral hepatitis, liver cirrhosis), chronic pancreatitis, chronic thyroiditis, phlegmosis (any origin) 'its cause and connective tissue matrix $ _ / weight Conditions (including cancer). The present invention relates to certain chemical compounds in the treatment of compulsive diseases. [Previous technology] In the developed and developed countries, some are related to scar formation and / or -6- (2) (2) 200302105 The occurrence of fibrosis-related diseases consumes a lot of resources. The costs of domestic and international public health plans to deal with the consequences of these diseases are also considerable. Therefore, it would be desirable to provide a method for treating or reducing the effects of diseases associated with scarring and / or fibrosis. The progression of certain diseases related to scar formation and / or fibrosis (such as atherosclerosis) may involve the accumulation / proliferation of smooth muscle cells (SMCs), which may cause extracellular matrix micromolecules (which (Mostly colloidal) becomes complicated. The process of atherosclerosis from thrombosis to myocardial infarction (MI) can lead to tissue damage, which can lead to scar tissue renewal and fibrous tissue formation. Although the normal wound repair process after tissue injury causes fibroblast proliferation, the process of fibroblast differentiation into muscle fibroblasts can be an early feature in the development of tissue fibrosis. The prolonged presence of myofibroblasts at the infarct site may also cause an imbalance between extracellular matrix proteins and proteases, which may worsen hypertrophic scars and wound formation. It would be desirable to provide a compound for the treatment of diseases associated with scarring and / or fibrosis, which compound is required to treat or at least alleviate these disease states. A composition for reducing scar formation is disclosed in EPO 93 0 069. Among these compositions, phosphodiesterase inhibitors are claimed to reduce scarring in wounds. However, the phosphodiesterase inhibitors described in this document are broad-spectrum inhibitors and are not specific to PDE 5. Therefore, the inhibitors of this patent do not have the same therapeutic effects as the compounds of the present invention, so they may be less advantageous.

Redondo '%: (Brit is h Journal of Pharmacology) 998, (3) (3) 200302105 124, M 5 5-1 4 62) illustrates a kind of atrial natriuretic peptide (ANP) and cyclic GMP phosphate The inhibitory effect of diesterase on collagen synthesis in adult cardiac fibroblasts. Two major subtypes of natriuretic peptide receptors have been identified, of which NPR-C is the largest, accounting for 70% of the natriuretic peptide receptor population in cardiac fibroblasts. The authors found that the PDE inhibitor, Zaprin st, itself had no effect on the regulation of cardiac fibroblast proliferation. Similarly, ANP itself has no effect on regulating the proliferation of cardiac fibroblasts. However, the combination of ANP and Sharprinster can indeed inhibit thymidine incorporation in a concentration-dependent manner within a limited concentration range, and the authors use this as an indirect analysis of DNA synthesis. The combination of C-ANF4_32 (NPR-C-specific analogue) and Sharprinster did not achieve this result repeatedly. The author states that another suspicion of this investigation is that Sharprinst is a specific PDE 5 inhibitor, however, in fact, the Sharplint system is recorded in other literature as a non-specific PDE Inhibitors (see, e.g., McMahon, et al. (Doc 18) and Kukoretz, et al. (Doc 3). It is also known that Sharplinster inhibits pdE 6 more than five times as strongly as PDE 5.

Duncan 'et al. (The FA SEB Journal) revealed an intravascular study conducted in the kidneys of normal rats, which found that connective tissue growth factor can mediate fibroblasts induced by transforming growth factor-P (TGF-yS) Collagen synthesis, while blocking CT GF synthesis or action in vivo, can reduce the formation of granulated tissue induced by TGF-by inhibiting collagen synthesis and fibroblast accumulation. cAMP can also inhibit collagen synthesis induced by CTGF itself ', but the report states that cGMP has no effect. This paper is related to (4) (4) 200302105

Redondo, et al. Contradicts the hypothesis that PDE 5 inhibits collagen production. Therefore, this technique cannot explain the role of cGMP in scar formation. Tissue repair after permanent damage involves a cascade of coordinated and overlapping overlapping phases. These reactions include inflammation, formation of granulated tissue, deposition and combination of extracellular matrix, and termination of the response. Peptide factors are involved in this process in different ways, and control platelet function, white blood cell tropism, cytokine synthesis, and angiogenesis and direct the progress of fibroblast phenotype, eventually forming premature scar tissue. Peptide factors control these processes by adjusting the ability of fibroblasts to proliferate, and quantitatively and qualitatively alter the sketching of their extracellular matrix components. One of the major regulators known to be involved in initiating a wound healing tandem response is TGF-cold. The literature suggests that nitric oxide can improve the healing rate of wounds. It is also known that cGMP PDE5 inhibitors can increase the intracellular concentration of cGMP derived from nitric oxide, thereby enhancing the effect of nitric oxide, which is related to the efficiency of treating sildenafil in male erectile dysfunction. Without wishing to be limited by any theory, it is generally believed that the effect of anti-scarring is related to the specific inhibition of PDE 5 at the appropriate stage in the wound healing cycle. This may occur with appropriate signals, such as smooth muscle relaxation mediated by NO. Other factors may also be involved. [Summary of the Invention] Surprisingly, we have therefore found that the administration of a PDE 5 inhibitor to a healing wound can reduce the formation of scar tissue. We observed in vivo that compared with normal heart tissue, -9- (5) (5) 200302105 showed excessive PDE5 protein in fibrosis of heart tissue. We have also determined that the PDE5 line is present in fibroblast subpopulations called myofibroblasts. In these cells, the expression of PDE5 is increased, so it may be involved in the pathophysiology of tissue fibrosis. In all tissues, the mechanisms that cause fibrosis are thought to be similar to each other, so fibrosis that occurs in the liver, kidneys, lungs, spine, and skin occurs in a similar manner. According to the present invention, the fibrosis of all these tissue types (and many other tissues) can be alleviated by inhibiting PDE5 to obtain significant curative effects. Although in the study of Sharplinster, data on non-selective PDE inhibition (such as those shown by Redondo, etc.) and the use of pentoxifylline (a weak and non-selective PDE inhibitor) have been obtained The idea that these agents can be used as anti-fibrotic agents has not been identified in the conventional arts, but selective PDE5 inhibition can be used as a basis to prevent or reduce scar formation. Indeed, with regard to the role of cGMP in scar formation (that is, the role of PDE 5), the point of view in the art of learning seems somewhat contradictory. According to a first aspect of the present invention, there is provided a method for reducing scar formation and / or treating fibrosis in a patient, which comprises treating the patient with an effective amount of a cGMP PDE5 inhibitor or a pharmaceutical composition thereof. A second aspect of the invention provides the use of a cGMP PDE5 inhibitor for the manufacture of a medicinal product for reducing scar formation and / or treating fibrosis. According to a third aspect of the present invention, there is provided the use of cGMP PDE5 inhibitor (6) (6) 200302105 for reducing scar formation and / or treating fibrosis. According to a fourth aspect of the present invention, there is provided a pharmaceutical pack comprising: a composition comprising a cGMP PDE5 inhibitor, instructions for use of the cgmP PDE5 inhibitor in reducing scar formation and / or treating fibrosis, and A container. In one embodiment of each of the above aspects, diseases related to scar formation and / or fibrosis that can be treated according to the present invention include: pulmonary fibrosis, atherosclerosis, cardiovascular disease, infection, trauma, surgery or Scar formation and / or fibrosis of the skin and / or cornea after thermal injury, scleroderma and its connective tissue disorders' fibrosis of the heart, chronic obstructive pulmonary disease, muscle fibrosis, renal fibrosis, chronic skin ulcers and Cleft scleroderma 'pulmonary fibrosis or any origin), post-operative and spontaneous adhesions, skin inflammation (including moss and related conditions), aging and all aging-related degeneration disorders (including aging of the skin), Liver fibrosis or any etiology (including viral and non-viral hepatitis, cirrhosis), chronic pancreatitis, chronic thyroiditis, calcinosis (any origin) 'its cause and accumulation of connective tissue matrix / Reshape the situation (including cancer). A therapeutic agent that selectively acts on cGMP PDE5 isoenzymes to improve or prevent scarring in tissues is not currently on the market. In the context of the present invention, a PDE5 inhibitor refers to any compound that is a potent and selective inhibitor of cGMP PDE5 isoenzyme. In order to achieve the purpose of the present invention, a PDE5 inhibitor must show a selectivity that preferentially inhibits PDE5 by at least 25 times, preferably at least 30 times. (7) (7) 200302105 Suitable for PDE5 in a pharmaceutical composition according to the present invention The inhibitor is a cGMP PDE5 inhibitor which will be described in detail below. It is particularly suitable for cGMP PDE5 inhibitors which are effective and selective here. Suitable cGMP PDE5 inhibitors for use in the present invention include: pyrazolo [4,3-d] pyrimidin-7-ones shown in EP-A-0463 7 5 6; disclosed in EP-A-0 5 26 Pyrazolo [4,3-d] pyrimidin-7-ones in 004; Pyrazolo [4,3-d] pyrimidinones disclosed in published international patent application W093 / 06 1 04; Revealed Isomerized pyrazolo [4,3-d] pyrimidin-4-ones in published international patent application WO93 / 07 1 49; quinazoles shown in published international patent application W093 / 1 2095 Porphyrin-4-ones; pyrido [3,2-d] pyrimidin-4-ones disclosed in published international patent application WO94 / 05 6 6 1 · disclosed in published international patent application W094 / Purine-6-ones in O04 5 3; pyrazolo [4,3-d] pyrimidin-7-ones disclosed in published international patent application WO 9 8/49 1 66; disclosed in Published international patent application W09 9/543 3 3 for pyrazolo [4,3-dp melidin-7-ones; disclosed in EP-A-0 9 95 7 5 1 for pyrazolo [4 , 3-d] chewing, pyridin-4-ones; disclosed in published international patent applications W 0 0 0/2 4 7 4 5 Pyridine and [4,3-d] pyrimidin-7-ones Class Pyrazolo [4,3-d] pyrimidin-4 -ones in e P-A-0 9 9 5 7 50; disclosed in published international patent application W 0 9 5/1 9 9 7 8 Compounds disclosed in published international patent application WO9 9/244 33 and compounds disclosed in published international patent application WO93 / 〇7124. Disclosed in published international patent applications W 0 0] / 2 7]] 2 Pyridine (8) (8) 200302105 Zolo [4,3-d] pyrimidin-7-ones; disclosed in published international patents Pyrazolo [4,3-d] pyrimidones in application WO 0 1/27 1 13; compounds disclosed in Ep · A-1 0927 19. Preferred type V phosphodiesterase inhibitors that can be used in the present invention include: 5- [2-ethoxy-5- (4-methyl-1-hexahydropyridinesulfonyl) phenyl] p-methyl _3-n-propyl_1,6-dihydro-711-pyrazolo [4,3- (1) pyrimidin_7-one (sildenafil), also known as 1 · [[3- ( 6,7-dihydro-1-methyl-7 · keto-3_propyl-1Η-pyrazolo [4,3-d] pyrimidin-5-yl) -4 · ethoxyphenyl] sulfonium Methyl] -4-methylhexahydropyridine (see EP-A-0463756); 5- (2-ethoxy-5-morpholinylacetamidophenyl-1-methyl-3 -n-propyl -1,6-dihydro-7: «-pyrazolo [4,3- (1] pyrimidin-7-one (see five? -Eight-0526004); 3-ethyl-5- [5- (4 -Ethylhexahydropyridine-1 -Sulfosulfanyl) -2-n-propoxyphenyl] -2- (pyridin-2-yl) methyl-2,6-dihydro-7H-pyrazolo [4,3-d] Pyrimidin-7-one (see WO 9 8/49 1 66); 3-ethyl-5- [5- (4-ethylhexahydropyridine-1 -ylsulfonyl) 2- (2-methoxyethoxy) pyridin-3-yl] -2- (pyridin-2-yl) methyl-2,6-dihydro-7H-pyrazolo [4,3-d] Pyrimidine-7-one (see W 0 9 9/5 4 3 3 3); (+) -3-ethyl-5- [5- (4-ethylhexahydropyridine-1 -ylsulfonyl) -2- (2-methoxy-1 (R) -methylethoxy) 氧基-3 -Methyl] -2 · methyl-2,6 -monohydro-7fluorene-pyrazolo [4,3-d] pyrimidin-7-one, also known as 3-ethyl-5- {5- [ 4-ethylhexahydropyridine-1 -ylsulfonyl) -2- ([(] R) -2-methoxy-1-methylethyl] oxy) pyridine-3 -yl 2. Methyl-2,6-dihydro-7H-pyrazolo [4,3-d] pyrimidine-7-amyl (see W 09 9/5 4 3 3 3); 200302105 Ο) 5- [2-ethoxy -5- (4-ethylhexahydropyridine-1-ylsulfonyl) pyridine_yl] -3-ethyl-2- [2-methoxyethyl] -2,6-dihydro-711 -Pyrazolo [4,3- (1) oral-coated-7-one 'is also known as 1- {6-ethoxy-5- [3-ethyl-6,7-monohydro-2- (2-Methoxyethyl) -7-keto-2H-pyrazolo [4,3-d] pyrimidin-5-yl] -3-pyridylsulfonyl 4-ethylhexahydropyridine D well ( See WO 01/271 13, Example 8);

5- [2-Iso-butoxy-5- (4-ethylhexahydropyrine-1-ylsulfonyl) pyridin-3-yl] -3 -ethyl-2- (1-methyl Hexahydropyridin-4-yl) -2,6-dihydro-7H-pyrazolo [4,3-d] pyrimidin-7-one (see WO 01/271 13 'Example 15); 5- [2-ethoxy-5- (4-ethylhexahydropyridine-butylsulfonyl) pyridinyl group] -3-ethyl-2-phenyl-2,6 · dihydro-7H- Pyrazolo [4,3-d] pyrimidin-7-one (see WO 01/271 13, Example 66); 5- (5-ethylfluorenyl-2-propoxy-3-pyridyl) -3 -Ethyl-2- (1-isopropyl-3-azetidinyl) -2,6-dihydro-7H-pyrazolo [4,3-d] pyrimidin-7-one (see WO 01/271 12 Example 124);

5- (5-ethylfluorenyl-2 · butoxy-3 · pyridyl) -3-ethyl-2- (1-ethyl-3-azetidinyl) -2,6-dihydro-71 ^- Pyrazolo [433- (1) pyrimidin-7-one (see ~ 0 01/271 12, Example 132); (6R, 12aR) -2,3,6,7,12,12a-hexahydro-2 -Methyl-6- (3,4-methylenedioxyphenyl) -pyracino [2,, 1 1: 6,1] pyrido [3,4-b] D 4-dione (IC-3 5 1) 'is the compound of Examples 7 8 and 95 in the published international patent application w 009 5/1 9 9 7 8; and Examples 1, 3, 7 And the compound in 8. 2- [2-Ethyl-5- (4-ethyl-hexahydro D to D-Dingyl fluorenyl) · phenyl] -5-methyl-7-propyl-3 Pyrene-imidazo [5 J-f] [], 2,4] trione-4-ketone (W-14- (10) 200302105 vardenafil), also known as 1-[[3-( 3,4-dihydro-5 -methyl-4-keto-7-propylimidazo [n-fj-as-trigen-2-yl) -4-ethoxyphenyl] sulfofluorene 4 -Ethylhexahydropyridine, which is the compound of Examples 20, 19, 337 and 336 in published international patent application W099 / 24433. Published in international patent application WO 93/0 7 1 24 Example 1 1 compound (EIS AI); and compounds 3 and 14 from Rotella DP, J. Med. Chem., 2000, 43, 127. Φ

Other types of cGMP PDE5 inhibitors that can be used with the present invention include: 4-ol-5- (pyridylmethylamino) -6- [3- (4-chlorophenyl) -propoxy] -3 (2H) Oral coaxing wakes up I, l- [4-[(l, 3-benzo-penta-5-ylmethyl) amino] -6-qi-2. Orally quinazolinyl] -4-hexahydro Pyridine-carboxylic acid, monosodium salt; (+)-cis-5,6 & 37,939,9 & -hexahydro-2- [4- (trifluoromethyl) -benzyl-5-methyl-cyclo Pentam-4,5] imidazole and [2,1-1 >] purha-4 (311) ketone; fluoro / raloxel (£ '11 ^ 21〇 (^ 11111); cis-2-hexyl -5-methyl-3,4,5,6 &, 7,8,9,9 & -octahydrocyclopenta [4,5] -imidazo [2, Ι-b] purin-4-one; 3 -Ethenyl-1- (2-chlorobenzyl) -2-propylindole-6-carboxylic acid ester; 3-ethenyl-1- (2-chlorobenzyl) -2-propylindole 6-carboxylic acid esters; 4-bromo-5- (3-pyridylmethylamino) -6- (3- (4-chlorophenyl) propoxy-3- (2H) dacrotonone; methyl-5 -(5-morpholinylacetamido-2-n-propoxyphenyl) -3-n-propyl-1,6-dihydro-7H-pyrazolo [4,3-d] pyrimidine-7 -Ketone; 1- [4-[(1,3-benzodifluoren-5-ylmethyl) amino] -6-chloro-2-oxazolinyl] -4-hexahydropicolinic acid, Monosodium salt; Pharmaprojects 4516 (Glax o Wellcome); Pharmaprojects 505] (Bayer);

Pharma projects 5 0 6 4 (K y o w a H a k k o; see W 0 9 6/2 6 9 4 0); -15- (11) (11) 200302105

Pharmaprojects 5 069 (Schering Plough); GF-1 96960 (Glaxo Wellcome); E-8010 and E-4010 (Eisai); Bay-38-3 04 5 & 3 8 -94 5 6 (Bayer) and Sch- 5 1 8 6 6. In order to avoid uncertainty, the PDE5 inhibitory compounds detailed above in the published reference patent specification described above specifically form part of the present disclosure and represent a part of the inventive subject matter of the present application. The suitability of any particular cGMP P.DE5 inhibitor can be easily assessed for its potency and selectivity using methods in the literature, and then evaluated for toxicity, absorption, metabolism, pharmacokinetics, etc. based on standard pharmaceutical practice Decide. A more suitable situation is that the cGMP PDE5 inhibitor has a 1C 50 of PDE5 of less than 100 nanomolar, more preferably less than 50 nanomolar, and more preferably less than 10 nanomolar. The IC50 of cGMP PDE5 inhibitors for PDE5 can be easily determined using established literature methods such as those described in EP0463756-B1 and EP0526004-A1. More suitably, the cGMP PDE5 inhibitor used in the present invention is a PDE5 enzyme. It is more appropriate to prefer PDE5 over PDE3, and it is more appropriate to prefer PDE5 over PDE3 and PDE4. More suitably, the selectivity ratio of the cGMP PDE5 inhibitor of the present invention to PDE5 to PDE3 (preferably the selectivity to PDE3 and PDE4) is greater than 25, preferably more than 3 G, and more preferably more than 00. The best inhibitors showed a selectivity ratio of PDE5 to PDE3 greater than 300. Those skilled in the art can easily determine the selectivity ratio. For PDE3 (12) (12) 200302105 and PD E4 enzyme I c 5 〇4 can use established literature methods, such as: s A Ballard, et al., Journal 〇 Urology, 1 998, v 〇l.159, pages The method described in 2164_2〗 71 can be easily determined. In order for the treatment to be effective, the compounds of the present invention are preferably orally bioavailable. Oral bioavailability refers to the proportion of orally administered drug that reaches the systemic circulation. The factors that determine the oral bioavailability of a drug are solubilization, membrane permeability, and metabolic stability. Usually, a screening tandem reaction is performed in vitro first, and then the in vivo technology is used to determine the oral bioavailability. Solubility (the case where the drug is dissolved by the aqueous content in the gastrointestinal tract (GIT)) can be achieved in a glass tube. Solubility experiments were performed to predict the GIT at a suitable pH. More suitably, the minimum solubility of the compound of the present invention is 50 µg / ml. Solubility can be determined by standard procedures in the art, such as those described in Adv. Drug Deliv. Rev. 23, 3-2 5, 1 997. Membrane permeability refers to the pathway of compounds through the cells of GIT. Lipophilicity To predict the key properties of this item, it is defined in a glass tube using LogD7.4 measurements using organic solvents and buffers. More suitably, the compound of the present invention has a LogD7.4 of -2 to +4, preferably -1 to +2. logD can be determined by standard procedures known in the art, such as: J. Ph a r m. Ph a r m a c ο 1. 19 9 0, 4 2:] 4 4. Cell monolayer analysis', such as ca co-2, can be used to essentially predict the membrane permeability allowed in the presence of outflow transporters (eg, para-glycoprotein), which is called caco-2 flux. More suitably, the flux of -17- (13) (13) 200302105 of the compound of the present invention is greater than SxlO ^ cms · 1, and preferably greater than SxlO ^ cms · 1. The caco flow rate can be determined by standard procedures in the art, such as those described in J. Pharm. Sci., 1 990, 79,595-600. Metabolic stability refers to the ability of GIT or the liver to metabolize compounds during absorption: the first pass-through effect. Analysis systems such as: microsomes, hepatocytes, etc. are predictors of metabolic tendency. It is more appropriate that the compounds of the examples show a metabolic stability of less than 0.5 in the analysis system equivalent to liver extraction. Examples of the analysis system and data manipulation are described in Curr. Opin. Drug Disc. Devel., 201, 4, 36-44, Drug Met. Disp., 2000, 28, 1 5 1 8- 1 523. The interaction of the above methods has further confirmed that the oral bioavailability of drugs in humans can be obtained by in vivo experiments performed in animals. In these studies, the compounds are administered separately or as a mixture via the oral route to determine absolute biological effectiveness. Absolute bioavailability can also be determined via the intravenous route (% absorbed). Examples of assessing oral bioavailability in animals can be found in Drug Met. Di sp., 2001, 29, 8 2-8 7; J. Med. Chem., 1 997, 4 0, 8 27-829, Drug Met. Disp., 1999, 27, 221-226. More suitably, the cGMP PDE5 inhibitor is sildenafil. The cGMP PDE5 inhibitor can be administered alone, but in human treatment, it is usually mixed with a suitable pharmaceutical excipient, diluent or carrier selected after referring to the intended route of administration and standard pharmaceutical methods. One's medication. For example: cGMP PDE5 inhibitors can be used for immediate-, delayed- (14) (14) 200302105, modified-, or controlled-release following medicinal forms for oral, oral or sublingual administration: lozenges , Capsules, pessaries, elixirs, solutions or suspensions. Such lozenges may contain excipients of the following groups: microcrystalline cellulose, lactose, sodium citrate, calcium carbonate, calcium phosphate dibasic salt and glycine, disintegrating agents such as: starch (preferably corn, potato , Or tapioca starch), sodium glycolate starch, croscarme (sodium croscarme), and some reconstituted silicates, and granulating linkers, such as polyvinylpyrrolidine Ketones, hydroxypropyl methylcellulose, hydroxypropyl cellulose, sucrose, gelatin and acacia. In addition, lubricants such as magnesium stearate, stearic acid, glyceryl lignoate, and talc may be included. A similar type of solid composition can also be used as the filling material in the capsule. In this regard, preferred excipients include: lactose, starch, cellulose, milk sugar or high molecular weight polyethylene glycols. In terms of water-soluble suspensions and / or tinctures, the cGMP PDE5 inhibitor of the present invention can be used with different sweeteners or flavoring agents, hueing substances or dyes, with emulsifying and / or suspending agents, and with diluents, Such as: water, ethanol, propylene glycol and glycerin, and combinations thereof. cGMP PDE5 inhibitors can also be administered parenterally, such as [[Intravenous, intra-arterial, intraperitoneal, intramuscular or subcutaneous route, or they can be administered by injection technology in such parenteral For administration, it is best to administer it in the form of a sterile aqueous solution (which may contain other substances such as sufficient salt or glucose to make it isotonic with blood). If necessary, this aqueous solution should be properly buffered (preferably adjusted to pH 値 between 3 and 9). Suitable parenteral formulations of -19- (15) (15) 200302105 can be easily prepared under sterile conditions by standard pharmaceutical techniques well known to those skilled in the art. The following dosages and other dosages in this article are for ordinary people weighing between 65 and 70 kg. Those skilled in the art can determine the dosage required for individuals whose weight is not within this range (such as children and the elderly). In this type of formulation, the dosage of the cGMP PDE5 inhibitor is based on its effectiveness, but can be It is expected to be in the range of 1 to 500 mg for up to three administrations per day. For oral and parenteral administration in human patients, the daily dose of cGMP PDE5 inhibitors is usually between 5 and 500 mg (in single or divided doses). In the case of sildenafil, the preferred dose is in the range of 10 to 100 mg (eg, 10, 25, 50 and 100 mg), which can be administered once or twice daily Or three times (preferably once). However, the exact dose will be determined by the clinician, and this dose will depend on the patient's age, weight and severity of symptoms. Therefore, for example, cGMP PDE5 inhibitor tablets or capsules may contain about 5 to 25 mg (eg, 10 to 100 milligrams) of the active compound, and may be administered in a single dose or divided into two parts over a period of time as appropriate. Or multiple doses. In any case, the clinician will determine the exact dosage that is most suitable for any individual patient, and this dosage will vary with the particular patient's age, weight, and response. The above dosages are just examples for general cases. Of course, there are individual cases where higher or lower dosages are used to achieve better results, and this is also within the scope of the present invention. cGMP PDE5 inhibitors can also be administered intranasally or by inhalation, and -20- (16) (16) 200302105 This can be conveniently used as a dry powder inhaler from a pressurized container, tub, sprayer, or sprayer Type, or atomized drop spray type, and use a suitable propellant, such as: dichlorodifluoromethane, trichlorofluoromethane, dichlorotetrafluoroethane, a hydrofluoroalkane, such as: 1,1J, 2- Tetrafluoroethane, or 1, I, 1,2,3,3,3-heptafluoropropane, carbon dioxide or other suitable gas delivery. In the case of a pressurized atomized drop, the dosage unit can be determined by providing a valve to deliver a calculated amount. The pressurized container, drum, sprayer, and diffuser may contain a solution or suspension of the cGMP PDE5 inhibitor, such as using a mixture of ethanol and a propellant as a solvent, and the solvent may additionally contain a lubricant. Such as sorbitan trioleate. Capsules and cartridges (made of, for example, gelatin) for use in inhalers or blowers can be formulated as a powder mixture containing cGMP PDE5 inhibitor and a suitable powder base such as lactose or starch. Nebulized drops or dry powders should preferably be arranged so that each calculated dose or "blow" contains cGMP PDE5 inhibitor from 1 to 50 mg for delivery to the patient. The total daily dose delivered in nebulized droplets will be between 1 and 50 mg, which may be administered to the patient as a single dose, or more often, in divided doses throughout the day. Alternatively, the cGMP PDE5 inhibitor may be administered as a suppository or vaginal suppository. The cGMP PDE5 inhibitor can be administered topically, and can be administered in the form of: gel, hydrogel, emulsion, solution, cream, ointment or dusting powder. cGMP PDE5 inhibitors can also be administered via the skin or percutaneous route, such as using a skin patch. -21-(17) (17) 200302105 For topical administration to the skin, cGMP PDE5 inhibitors can be formulated into suitable ointments, where the inhibitor is suspended or dissolved in, for example, one or more of the following substances In the mixture ... Mineral oil, liquid petroleum jelly, white petroleum jelly, propylene glycol, polyethylene oxide polyoxypropylene compound, emulsified wax and water. Alternatively, it can be formulated into a suitable emulsion or cream, wherein the inhibitor is suspended or dissolved in, for example, a mixture of one or more of the following: mineral oil, sorbitan monostearate, Polyethylene glycol, liquid paraffin, polysorbate 60, paraffin esters of cetyl esters, cetylaryl alcohol, 2-octyldodecanol, benzyl alcohol and water. cGMP PDE5 inhibitors can also be used in combination with cyclodextrin. It is known that cyclodextrin can form contained and non-included complexes with drug molecules. Forming a drug-cyclodextrin complex can modify the solubility, rate of dissolution, bioavailability, and / or stability of a drug molecule. In general, drug-cyclodextrin complexes can be used in most dosage forms and routes of administration. Cyclodextrins can be used as auxiliary additives, such as: as a carrier, diluent or co-solvent, instead of directly compounding with drugs. α-, Θ- and r-cyclodextrin are the most commonly used, and suitable examples are described in WO-A-91 / 11172, WO-A-941 / 02518, and WO-A-98 / 55148. In general, in humans, cGMP PDE5 inhibitors are better administered orally, and are the most convenient. When the user has a swallowing disorder or the drug is malabsorbed after oral administration, the drug can be administered parenterally, sublingually, or orally. The cGMP PDE5 inhibitor of the present invention can also be used with one or more of the following (18) 200302105 i) a-adrenergic receptor antagonist compounds, also known as α-adrenoceptor, or α-adrenoceptor Body, or alpha-blockers. Compounds suitable for use herein include alpha-adrenergic stress as described in PCT Application WO " / 3069697, published on June 14, 1998

Receptors, of which the disclosure of α-adrenergic receptors is reference material for this article, including: selective ai-adrenergic receptors or α 2-adrenergic receptors , And non-selective adrenergic receptors, suitable α! -Adrenergic receptors include: phenazoline, phenazoline mesylate, clopitridone, oufa Alfuzosin, indipamide, naftopidil, tamsulosin, dapiprazole 'phenoxybenzamine, idazoxan, efaraxan , Yohimbine, Rovewood Bioassay, Recordati 1 5/273 9, SNAP 1 069 > SNAP 5 0 8 9, RS17053, SL89.0591, doxazosin, special Terazosin, abanoquil, and erbium; and blockers from US 6,037,346 [March 14, 2000]: dibennarnine, benzazoline, Trimethoxyoxazoline; an alpha-adrenergic receptor as described in the following US patent applications: 4,1 8 8,3 90; 4,026,894; 3,511,836; 4,3 15,007; 3,527,761; 3,997,666; 2,503,059; 4,703,063; 3,381,009; 4,252,721 and 2,5 9 9,0 0, each of which is a reference material; α 2 -adrenergic receptors include: clonidine , Papaverine, papaverine hydrochloride, can be present at will-a kind of cariton (nic) agent, such as: pir X amine; Π) NO-donor (N 0-agonist) compound. Suitable for use here -23- (19) 200302105

NO-supplier compounds include: organic nitrates, such as: mono-, di-, or tri-nitrates' or organic nitrates, including: glyceryl trinitrate (also known as nitroglycerin), 5-- Isosorbide dinitrate, isosorbide dinitrate, erythritol tetranitrate, erythritol tetranitrate, sodium nitrocyanate (SNP), 3-morpholinosydnonimine, S- Nitroso-N-acetamido penicillamine (SNAP), S-nitroso-N-glutamine sulfur (SNO-GLU), N-hydroxy-L-arginine, amyl nitrate, Lins Ketone amine (iinsidomine), linsone amine chloride hydrate, (SIN-1) S-nitroso-N-cysteine, diazenium diolates, (NONOates), 1, 5 -Glutaric nitrate, L-arginine, human tincture, jujube, morphone, Re-2047, nitrosed maxislyte derivatives, such as published PCT application WO 00 NMI-67 8- 1 1 and NMI-937 described in 1 2 07 5

iii) a vasodilator. Vasodilators suitable for use herein include: nimodepine, pinacidil, cyclomandelate, phenoxypropanolamine, chloroprumazine, haloperidol, Rec 15/2739, lopertriazone, pentoxifylline; iv) thromboxane A 2 agonist; v) NO-synthetic enzyme substrates, such as: arginine; vi) calcium channel blockers, such as: Amlodipine; vii) steroidal or non-steroidal anti-inflammatory agents; viii) parent metalloproteinase inhibitors (ΜΜΡ), especially ΜΜ P-3, Μ Μ Ρ-I 2 and Μ Ρ Ρ−3; ix) urinary hormone cytosinogen activator inhibitor (UPA); -24- (20) 200302105 x) pcp inhibition Qi: and xi) PDE4 inhibits Qi U. Agents particularly suitable for use in combination with the PDE5 inhibitor of the present invention to treat wounds include: PCP inhibitors, such as those described in WO 0 1/4790 1, GB0108097.7, PCT / IB01 / 02360, and GB0108102.5.

More suitably, the MMP inhibitor is a MMP-3 and / or MMP-13 inhibitor, such as: specifically and generally disclosed in WO 99/3 5 1 24, EP 9 3 1 7 8 8, WO 99/29667 or WO 00/74681. Particularly preferred MMP inhibitors are those disclosed in the examples of WO 99/35124, EP931788, WO 99/29667 and WO 00/74681.

More suitably, the uPA inhibitor is selected from those specifically and generally disclosed in WO 99/20608, EP1044967 or WO 00/05214. Particularly preferred uPA inhibitors are those disclosed in the examples of WO 99/20608, EP1044967 and WO 0/0 5 2 14.

All references to treatment in this article include curative, palliative, and prophylactic treatments. The application of the present invention is illustrated by the following figures, among which: guinea-human multiple strains of anti-blood pupa are produced in the rabbit, and are based on

Fawcett, et al. (Proc Natl Acad Sci USA 2000; 97: 3702 · 3 7 0 7) method for the affinity of LIP-1 [MERAGP SFGQ QR] peptide (which is equivalent to the amino acid residue M2 of human PDE5A1) Sexual purification. LIP-] is specific to PDE5 A1. 4 micron-sized formalin-fixed, paraffin-embedded tissue was cut and cut into 25- (21) (21) 200302105 pieces and placed in APE S (3-aminopropyltriethoxysilane) ) On a glass slide, and dried at 6 ° C for 1 hour. The sections were then dewaxed and rehydrated ', and then they were incubated at 37 ° C at 0.1% trypsin (in (^. /. Cl) In calcium (pH 7-6) [Proteolytic antigen retrieval] in [pH7-6] for 8 minutes. After briefly washing with water, it was placed in 9 ml of H2O2 aqueous solution (distilled water was added to make the total volume Incubate for 10 minutes in 10 (ml) to block the activity of endogenous peroxidase. The sections were rinsed in tap water and then transferred to PBS. The excess buffer solution on the slide was removed, and The test sections were incubated for 1 hour at room temperature in LIP-1 antibody (diluted in PBS at a ratio of 1: 600). The primary antibody was removed as a negative control group. The positive control tissue used was human cavernous body .Immune detection uses 3-amino-9-ethylcarbazole (3 AEC) as matrix chromogen (red / brown staining), using DAKO rabbit to imagine TM system (DAKO Rabbit Envision TM system). Figure 1 illustrates reactive but uninflamed skin sections at the edge of the skin wound. The expression of the positive staining lines of smooth muscle cells and negative fibroblasts in the middle layer of blood vessels of the microvenous PDE5 manifestations in healing wounds. Hyperplasia but intact squamous epithelium 1 is negative. The underlying dermis contains mature scar tissue with small and large micro veins 2. Note that in the middle layer of blood vessels Positive dark staining of the smooth muscle cells (original magnification X] 〇). Figure 2 shows the stone section taken from the edge of the ulcer (left) and intact epithelium (right) during the 4th day of healing. Similarly, positive staining of smooth muscle cells (right) in the middle layer of blood vessels of the venules and spindle (22) (22) 200302105 cells (myofibroblasts) in the bottom of the ulcer (left) indicates the expression of PDE5. Hyperplasia but Intact squamous epithelium (right) and necrotic inflammatory exudate 3 were negative. Note the smooth muscle cells in the middle layer of blood vessels in the microvenous 4 and the spindle cells in the bottom of the ulcer 5. Positive dark staining (original magnification x20). Figure 3 is a paraffin section taken from the bottom of a healed ulcer, in which immature scar tissue bundles have replaced normal skin structures. Similarly, some spindle cells ( Myofibroblasts (8) and the positive staining of certain vascular structures indicate the performance of PDE5 (original magnification X2.) Figure 4 is a higher magnification image of the skin litter section of Figure 3. This slice system Taken from the bottom of the healed ulcer, the immature scar tissue bundle has replaced the normal skin structure. The positive staining of some spindle cells (myofibroblasts) (9) and some microvessels (10) with a thin vascular middle layer indicates the performance of PDE5 (original magnification X40). Figure 5 is a higher magnification image of Figure 4 showing a section taken from the bottom of a healed ulcer, in which immature scar tissue bundles have replaced normal skin structures. In the section, some spindle cells (myofibroblasts) (11) were positively stained in acellular collagen. Some of these spindle cells have a patchy distribution of immunolocalization in the cytoplasm. The positive staining of middle smooth muscle cells in the small arteries 02) shows the performance of PDE5. Negative staining of the superficially arranged endothelial cells (13) indicates no P D E 5 (original magnification X 60). Figure 6 of the rabbit is also a higher magnification image of Figure 4, which shows a slice taken from the bottom of the healed ulcer, which is tied to a relatively immature scar tissue area. Similarly, the positive staining of certain spindle cells (Isofibroblasts -27- (23) (23) 200302105) (i4) and middle smooth muscle cells in the arteriolar (central) (15) represents PDE 5. Some of these spindle cells have a patchy distribution in the cytoplasm. (Original magnification x60). [Embodiment] The following formulation examples are only for illustration, and are not intended to limit the scope of the present invention. Active ingredient refers to a cGMP PDE5 inhibitor. Formulation 1: Use the following ingredients to prepare lozenges; blend sildenafil citrate (50 mg) with cellulose (microcrystalline form), silicon dioxide, stearic acid (smoked), and The mixture is compressed into a lozenge. Formula 2: The active ingredient (100 mg) is combined with isotonic physiological saline (1,000 ml) to prepare an intravenous formulation. Formulation 3: A topical formula can pass up to 2 weight. / 〇 The active ingredient is prepared by combining a suitable excipient, which can be a mixture of one or more of the following: mineral oil, liquid petrolatum, white petrolatum, propylene glycol, polyethylene oxide polyoxypropylene compound, emulsification Wax and water. -28- (24) (24) 200302105 [Schematic description] The first image is a microscopic image of a paraffin section of the skin magnified 10 times; the second image is a microscopic image of a paraffin section of the skin magnified 20 times; Figure 3 is a microscopic image of a paraffin section of skin magnified at 20 times; Figure 4 is a microscopic image of a paraffin section of skin magnified at 40 times; Figure 5 is a micrograph of a paraffin section of skin magnified at 60 times; And Figure 6 is a micrograph of a paraffin section of the skin at a magnification of 60 times. • Comparison of main components Table 1: Squamous epithelium 2: Microvenous 3: Necrotic inflammatory exudate 4: Microvenous 5: Ulcer 8: Spindle cells (myofibroblasts) 9: Spindle cells (myofibroblasts) ® 10: Microvessels 1 1: spindle cells (myofibroblasts) 1 2: arterioles 1 3: endothelial cells 1 4: clock cells (myofibroblasts)] 5: arterioles-29-

Claims (1)

(1) (1) 200302105 Patent application scope 1. A pharmaceutical composition for reducing scar formation and / or treating fibrosis in a patient, comprising a cGMP PDE5 inhibitor in an effective amount. 2. The pharmaceutical composition according to item 1 of the patent application, wherein the disease related to scar formation and / or fibrosis is selected from the following groups: pulmonary fibrosis, atherosclerosis, cardiovascular disease, infection, trauma, Scar formation and / or fibrosis of the skin and / or cornea after surgery or thermal injury, scleroderma and other connective tissue disorders, fibrosis of the heart, chronic obstructive pulmonary disease, muscle fibrosis, renal fibrosis, chronic skin ulcers And lip scleroderma, pulmonary fibrosis or any origin), postoperative and spontaneous adhesions, skin inflammation (including moss and related conditions), aging and all aging-related degeneration disorders (including skin aging), Liver fibrosis or any etiology (including viral and non-viral hepatitis, cirrhosis), chronic pancreatitis, chronic thyroiditis, calcinosis (any origin), its cause and accumulation of connective tissue matrix / Reshape the situation (including cancer). 3. Use of a cGMP PDE5 inhibitor for the manufacture of a medicinal product for reducing scar formation and / or treating fibrosis. 4. For the application in the scope of patent application item 3, wherein the disease related to scar formation and / or fibrosis is selected from the following groups: pulmonary fibrosis, atherosclerosis, cardiovascular disease, infection, trauma, surgery or Scar formation and / or fibrosis of the skin and / or cornea after thermal injury, scleroderma and other connective tissue disorders, fibrosis of the heart, chronic obstructive pulmonary disease -30- (2) (2) 200302105, muscle fibrosis , Renal fibrosis, chronic skin ulcers and scleroderma of the lips (pulmonary fibrosis or any origin), postoperative and spontaneous adhesions, skin inflammatory conditions (including moss and related conditions), aging and all related to aging Degenerative disorders (including skin aging), liver fibrosis or any etiology (including viral and non-viral hepatitis, cirrhosis), chronic pancreatitis, chronic thyroiditis, calcinosis (any origin), its causes Conditions related to the accumulation / remodeling of connective tissue matrix (including cancer). 5. The pharmaceutical composition or use according to any one of claims 1 to 4, wherein the inhibitor is administered orally or topically. 6. The pharmaceutical composition or use according to any one of items 1 to 4 of the patent application, wherein the inhibitor has an IC50 of less than 100 nanomolar. 7. The pharmaceutical composition or use as described in item 6 of the scope of the patent application, wherein the selective ratio of the inhibitor is more than 1,000. 8. The pharmaceutical composition or use according to any one of claims 1 to 4, wherein the inhibitor is sildenafil. 9. The pharmaceutical group, product or use in item 8 of the scope of patent application, wherein the daily dosage is 5 to 500 mg. 10. The pharmaceutical composition or use as described in item 9 of the scope of patent application, wherein the daily dosage is 10 to 100 mg. 11. Use of an eGMP PDE5 inhibitor in combination with a PcP and / or PDE4 inhibitor in the manufacture of a medicinal product for reducing scar formation and / or treating fibrosis. ] 2. An academy pack comprising: a drug (3) (3) 200302105 containing a PDE5 inhibitor, instructions for use of the composition in reducing scar formation and / or treating fibrosis, and A container. 13. A composition of a PDE5 inhibitor plus a PCP inhibitor and / or a PDE4 inhibitor (uP A). -32-