TWI469771B - System for providing controlled release delivery of drugs for treatment of neointimal hyperplasia and pharmaceutical composition for reducing neointimal hyperplasia - Google Patents

Description

A system for controlling release delivery of a drug for treating neovascular intimal hyperplasia and a pharmaceutical composition for reducing neovascular intimal hyperplasia

The present invention relates to novel compounds having immunomodulatory activity and/or anti-restenosis activity and synthetic intermediates for the preparation of such novel compounds, and in particular to macrolide immunomodulators. More specifically, the present invention relates to semi-synthetic analogs of rapamycin, methods of preparing the same, pharmaceutical compositions containing the same, and methods of treatment using the same.

Since the introduction of organ transplantation and immunomodulation, the broad use of the compound cyclosporine (cyclosporin A) has been found and has led to a significant increase in the success rate of the transplantation procedure. Recently, some species of macrocyclic compounds having potent immunomodulatory activity have been discovered. European Patent Application No. 184,162, issued to Okuhara et al., issued June 11, 1986, discloses a large number of macrocyclic compounds isolated from the genus Streptomyces , which are isolated from a strain of S. tsukubaensis . Immunosuppressant FK-506 (which is a 23-membered macrolide).

Other related natural products (such as FR-900520 and FR-900523, which differ from FK-506 in that their alkyl substituents on C-21) have been isolated from S. hygroscopicus yakushimnaensis . Another analog, FR-900525 (produced by Streptomyces faecalis), differs from FK-506 in that it replaces the hexahydronicotinic acid moiety with a proline group. Unsatisfactory negative effects associated with cyclosporine and FK-506, such as nephrotoxicity, have led to the continual search for immunosuppressive compounds with improved efficiency and safety, including locally effective but systemically ineffective immunosuppressive agents (US Patent No. 5,457,111).

Rapamycin is a macrocyclic triene antibiotic produced by Streptomyces hygroscopicus and found to have antifungal activity in vitro and in vivo, especially against Candida albicans (C. Vezina et al., J.Antibiot 1975,28,721;. SNSehgal et al J.Antibiot 1975,28,727;. HABaker et al. J. Antibiot 1978,31,539;. U.S. Pat. No. 3,929,992; and U.S. Patent No. 3,993,749).

Rapamycin, alone (U.S. Patent No. 4,885,171) or in combination with picibanil (U.S. Patent No. 4,401,653), has been shown to have anticancer activity. In 1977, rapamycin was also shown to be effective as an immunosuppressant in the experimental allergic encephalomyelitis model (model for multiple sclerosis) and the adjuvant arthritis model (model for rheumatoid arthritis). And showed its effective inhibition of the formation of IgE-like antibodies (R. Martel et al, Can. J. Physiol. Pharmacol . , 1977, 55, 48).

The immunosuppressive effect of rapamycin has also been disclosed in FASEB, 1989, 3, 3411 because it prolongs the survival of organ transplants in tissue incompatible rodents (R. Morris, Med. Sri. Res. , 1989, 17 , 609). The ability of rapamycin to inhibit T cell activation is revealed by M. Strauch ( FASEB, 1989, 3, 3411). These and other biological effects of rapamycin have been previously reviewed in Transplantation Reviews, 1992, 6 , 39-87. Rapamycin has been shown to reduce neovascular intimal hyperplasia in animal models and reduce the rate of restenosis in humans. It has been publicly shown that rapamycin also exhibits evidence of an anti-inflammatory effect, which supports its selection as a drug of choice for the treatment of rheumatoid arthritis. Since cytomycin and its analogs have been proposed to prevent restenosis because cell proliferation and inflammatory balloon angioplasty and stent placement are thought to form an inducing factor for restenosis.

Monoester and diester derivatives of rapamycin (esterification at positions 31 and 42) have been shown to be useful as antifungal agents (U.S. Patent No. 4,316,885) and water-soluble prodrugs of rapamycin (US Patent) No. 4, 650, 803).

The fermentation and purification of rapamycin and 30-desmethoxyrapamycin have been described in the literature (C. Vezina et al . J. Antibiot. (Tokyo), 1975, 28 (10), 721; SNSehgal et al. , J. Antibiot. (Tokyo), 1975, 28(10), 727; 1983, 36(4), 351; NL Paiva et al. J. Natural Products, 1991, 54 (1), 167-177).

A number of chemical modifications to rapamycin have been attempted. These include the preparation of monoester and diester derivatives of rapamycin (WO 92/05179), 27-oxime of rapamycin (EP0 467606), and 42-side oxy analogues of rapamycin (USA) Patent No. 5,023,262), bicyclo rapamycin (U.S. Patent No. 5,120,725), rapamycin dimer (U.S. Patent No. 5,120,727), rapamycin decyl ether (U.S. Patent No. 5,120,842) and Aryl sulfonate and amino sulfonate (U.S. Patent No. 5,177,203). Recently, rapamycin has been synthesized in its naturally occurring enantiomeric form (KCNicolaou et al, J. Am. Chem. Soc, 1993, 115, 4419-4420; D. Romo et al , J. Am. Chem. Soc , 1993, 115, 7906-7907; C. Hayward, J. Am. Chem. Soc, 1993, 115, 9345-9346).

Rapamycin such as FK-506 is known to bind FKBP-12 (Siekierka, JJ; Hung, SHY; Poe, M.; Lin, CS; Sigal, NH Nature, 1989, 341, 755-757; Harding, MW; Galat, A.; Uehling, DE; Schreiber, SL Nature 1989, 341, 758-760; Dumont, FJ; Melino, MR; Staruch, MJ; Koprak, SL; Fischer, PA; Sigal, NHJ Immunol. 1990, 144, 1418-1424; Bierer, BE; Schreiber, SL; Burakoff, SJ Eur. J. Immunol. 1991, 21, 439-445; Fretz, H.; Albers, MW; Galat, A.; Standaert, RF; Lane, WS; Burakoff , SJ; Bierer, BE; Schreiber, SL J. Am. Chem. Soc. 1991, 113, 1409-1411). It has also been shown that the rapamycin/FKBP-12 complex binds to another protein, m-TOR, which is different from calcineurin (FK-506/FKBP-12 complex inhibited protein) (Brown, EJ; Albers, MW; Shin, TB; Ichikawa, K; Keith, CT; Lane, WS; Schreiber, SL Nature 1994, 369, 756-758; Sabatini, DM; Erdjument-Bromage, H.; Lui, M.; P.; Snyder, SH Cell, 1994 , 78 , 35-43).

Other drugs have been used to combat unwanted cell proliferation. An illustrative example of such drugs is paclitaxel. Pacific paclitaxel, which is a miscellaneous alkaloid extracted from Pacific Yew and Taxus brevifolia , is a key cytoskeletal component in cell division (tubulin, the basic component of microtubules) ), thus preventing cell proliferation (Miller and Ojima, 2001).

In the 1970s, Andreas Gruntzig developed percutaneous transluminal coronary angioplasty (PTCA). The first canine coronary artery dilatation was implemented on September 24, 1975; the following year, the study showing the use of PTCA was presented at the annual meeting of the American Heart Association. Shortly thereafter, the first human patient was studied in Zurich (Switzerland), followed by the first American human patient in San Francisco and New York. Regarding the treatment of patients with obstructive coronary artery disease, although this procedure changes the implementation of interventional cardiology, this procedure does not provide a long-term solution. The patient receives only temporary relief of chest pain associated with vascular occlusion; routine procedures are often required. Determining the presence of restenosis lesions severely limits the use of this novel procedure. In the late 1980s, stents were introduced to keep blood vessels open after angioplasty. Stenting involves 90% of the angioplasty performed today. The rate of restenosis was 30-50% change in patients treated with balloon angioplasty prior to introduction of the stent. The introduction of stenting resulted in a further improvement in the outcome, with a restenosis rate of 15-30%. After stenting, restenosis damage is mainly caused by neointimal hyperplasia. Neovascular intimal hyperplasia and atherosclerotic disease are significantly different in time course and histopathological appearance. Restenosis is the healing process of a damaged coronary artery wall in which neovascular intimal tissue significantly impinges on the lumen of the blood vessel. Vascular radiotherapy appears to be effective in preventing under-stent restenosis. However, radiation has practical and cost-limiting limitations, and there are questions about safety and durability.

By reducing restenosis by at least 50%, the primary efforts made by the interventional device population to manufacture and evaluate drug elution stents have reached initial goals. However, there remains a need for improved topical drug delivery devices, such as drug-impregnated polymer coated stents, which provide a safe and effective tool for the prevention and treatment of restenosis. For example, two commercially available single drug-dissolving stents reduce restenosis and improve patient outcome, but they do not eliminate restenosis or avoid adverse safety issues. Patients and especially at risk (including diabetic patients with small blood vessels and patients with acute coronary syndrome) may benefit from topical drug delivery devices (including stents with improved capabilities).

Drug delivery devices comprising a combination of drugs are known. However, this technique does not appear to teach a particularly effective combination of drugs for topical administration (e.g., from stent dissociation). For example and as discussed further below, Falotico teaches an EVA-PBMA polymer coated stent comprising a combination of rapamycin/dexamethasone that reduces neovascular area, percentage of stenosis, and inflammation. The score is far from being effective in delivering a single rapamycin or dexamethasone alone.

In one aspect of an embodiment of the present invention, a compound represented by the following structural formula is disclosed:

Or a pharmaceutically acceptable salt or prodrug thereof.

In another aspect of the invention, the invention relates to a system for providing controlled release delivery of a medicament for treating or inhibiting neovascular intimal hyperplasia in a vascular lumen, comprising a therapeutic amount of a first medicament and a therapeutic amount a composition of a second drug, wherein the first drug comprises at least one olimus drug and salts, esters, prodrugs and derivatives thereof; and wherein the first drug is therapeutically effective and in a therapeutic amount of the second drug The activity of supplementing the second drug is present, and wherein the second drug is therapeutically effective and the activity of the first drug is supplemented in the presence of a therapeutic amount of the first drug.

In another aspect, the invention relates to a pharmaceutical composition for topical administration for reducing neovascular intimal hyperplasia comprising at least one Moss drug and at least one glucocorticol, wherein the Moss drug and the glucocorticol are at about 10 : A ratio between 1 and about 1:10.

In one aspect, embodiments of the invention are directed to a drug delivery system having a support structure comprising at least one pharmaceutically acceptable carrier or excipient and having zotarolimus and dexamethasone A therapeutic composition of (dexamethasone) or a derivative, prodrug or salt thereof, wherein the formation of neovascular intimal hyperplasia is reduced when the system is implanted into the lumen of the blood vessel of the subject as compared to the control system. The drug delivery system can include a stent and can include a third or more drug or other therapeutic substance (including biological agents). Other therapeutic substances include, but are not limited to, anti-proliferative agents, anti-platelet agents, steroid and non-steroidal anti-inflammatory drugs, anti-thrombotic agents, and thrombolytic agents. The subject can be a mammal, including but not limited to humans and pigs.

Another object of an embodiment of the present invention is to provide a synthetic method for preparing such compounds from the starting materials obtained by fermentation and a chemical intermediate suitable for such synthetic methods.

A further object of an embodiment of the present invention is to provide a pharmaceutical composition comprising at least one of the above compounds as an active ingredient.

Another object of embodiments of the present invention is to provide a method of treating various disease conditions including restenosis, post-transplant tissue rejection, immune and autoimmune dysfunction, fungal growth, and cancer.

In another aspect of an embodiment of the invention, a medical device comprising a support structure having a coating on at least a portion of its surface is provided, the coating comprising a therapeutic substance (such as a drug). Support structures for medical devices suitable for use in the present invention include, but are not limited to, coronary stents, peripheral stents, catheters, arterio-venous grafts, bypass grafts, and drug delivery balloons (including angioplasty) used in vascular structures Balloon). Drugs suitable for use in the present invention include, but are not limited to, Or a pharmaceutically acceptable salt or prodrug thereof, including Or a pharmaceutically acceptable salt or prodrug thereof (hereinafter referred to as quetiamus and A-179578), and Or a pharmaceutically acceptable salt or prodrug thereof; Or a pharmaceutically acceptable salt or prodrug thereof (hereinafter referred to as A-94507).

In another aspect, embodiments of the invention pertain to safe and effective local delivery of a pharmaceutical combination comprising quetiamus and dexamethasone. The combination of drugs provides improved safety and efficiency compared to single drug-eluting stents, including those found in current clinical practice (Taxus® and Cypher®).

In another aspect, embodiments of the present invention provide methods and devices to safely and effectively further reduce neovascular intimal area, neovascular intima thickness, and percentage of stenotic area associated with a local delivery device that delivers a drug combination. In particular, no combination of quetiamus/dexamethasone was observed to adversely affect the activity of each drug. Instead, separate the stent with a single drug (such as Taxus) And Cypher In comparison, the combination proves an improvement in safety and efficiency.

Coatings suitable for use in embodiments of the invention include, but are not limited to, polymeric coatings comprising any polymeric material in which the therapeutic agent is effectively dispersed. The coating can be hydrophilic, hydrophobic, biodegradable or biodegradable. This medical device releases the drug at an appropriate rate to effectively reduce restenosis in the vascular structure. It is expected that direct coronary artery delivery of a drug comprising quetiamus will reduce the rate of restenosis to a level of less than about 15%.

Definition of Terms

The term "related to" as used herein refers to a compound that can be in a variety of forms including, but not limited to, mixed, unmixed, microspheres, mixed with a coating, and mixed with a support structure. Those skilled in the art are aware of changes in the interaction between drugs, coatings/drugs, and drug/coating/support structures.

The term "supplement" as used herein refers to the behavior exhibited by at least two drugs in a combination wherein each of its pharmaceutically active agents benefits from a combination, for example, has a potentiating activity; for example, has separate but beneficial activities, such activity Helps all of the desired pharmacological effects in mammals; and wherein the combination drugs do not actively reduce each other's biological activity.

The term "prodrug" as used herein refers to a compound that is rapidly converted to the parent compound of the above formula by hydrolysis in the blood, for example. A detailed discussion by T. Higuchi and V. Stella, "Pro-drugs as Novel Delivery systems," in the 14th volume of the ACSSymposium Series and in Edward B. Roche, ed., "Bioreversible Carriers in Drug Design," A merican Pharmaceutical Provided in Association and Pergamon Press, 1987, both incorporated herein by reference.

The term "pharmaceutically acceptable prodrug" as used herein refers to a prodrug of a compound of the present invention which is suitable for contacting humans and lower mammals within the scope of sound medical judgment. Inappropriate toxicity, irritation, and allergic reactions are proportional to the reasonable benefit/hazard ratio and are effective for their intended use. Other examples include pharmaceutically acceptable prodrugs derived from C-31 hydroxyl groups.

R=R ¹ C(O)R ² R ³ , R ¹ C(S)R ² R ³ ; wherein R ¹ =O, S; R ² = none, O, N, S, various alkyl groups, alkenyl groups, Alkynyl, heterocyclic, aryl; R ³ = none, various alkyl, alkenyl, alkynyl, heterocyclyl, aryl; alkyl, alkenyl, alkynyl, heterocyclyl, aryl may be substituted Or unsubstituted.

The term "prodrug ester" as used herein refers to any of a number of ester-forming groups that are hydrolyzed under physiological conditions. Examples of the prodrug ester group include etidinyl, propyl sulfonyl, trimethylethenyl, pentyloxymethyl, ethoxymethyl, decyl, methoxymethyl, dihydroindenyl and Similar groups, as well as ester groups derived from the coupling of a natural or non-naturally occurring amino acid with a C-31 hydroxyl group of a compound of the present invention.

The term "support structure" means a framework that can include or support a pharmaceutically acceptable carrier or excipient, which can include one or more therapeutic agents or substances (eg, one or more drugs and/or Or other compound). The support structure is typically formed from a metal or polymeric material. Suitable support structures that can be formed from a polymeric material, including a biodegradable polymer, which can include a therapeutic agent or substance, include, but are not limited to, those disclosed in U.S. Patent Nos. 6,413,272 and 5,527,337, each incorporated herein by reference. They support the structure.

"Subject" means a vertebrate, including but not limited to mammals, including monkeys, dogs, cats, rabbits, cows, pigs, goats, sheep, horses, rats, mice, guinea pigs, and humans.

"Therapeutic substance" means any substance that exerts a beneficial effect on a subject when appropriately administered to a subject at an appropriate dose.

Example

In one embodiment of the invention, the compound of the formula

In another embodiment of the invention, the compound of the formula

treatment method

Compounds of the invention including, but not limited to, the compounds illustrated in the Examples have immunomodulatory activity in mammals, including humans. As immunosuppressive agents, the compounds of the embodiments of the present invention are useful for the treatment and prevention of immunomodulatory diseases, including diseases or tissues (including heart, kidney, liver, bone marrow, skin, cornea, lung, pancreas, small intestine (intestinum). Transplantation resistance of tenue), limbs, muscles, nerves, duodenum, small-bowel, islet cells and their analogues; graft-versus-host disease caused by bone marrow transplantation; including rheumatoid arthritis, whole body Autoimmune diseases of lupus erythematosus, Hashimoto thyroiditis, multiple sclerosis, myasthenia gravis, type I diabetes, uveitis, allergic encephalomyelitis, spheroid nephritis and the like. Other uses include the treatment and prevention of skin manifestations of inflammatory and hyperproliferative skin diseases and immune-regulating diseases, including psoriasis, atopic dermatitis, contact dermatitis and other eczema dermatitis, seborrheic dermatitis, lichenitis, Acne, bullous pemphigoid, bullous epidermis, rubella, angioedema, vasculitis, erythema, subcutaneous eosinophilia, lupus erythematosus, acne and alopecia areata; including dry keratoconjunctivitis, spring Conjunctivitis, uveitis associated with Behcet's disease, keratitis, herpetic keratitis, keratoconus, corneal epithelial dystrophy, corneal leukoplakia, and various ocular diseases of eye pemphigus (autologous regulation and others). In addition, including asthma (such as bronchial asthma, allergic asthma, endogenous asthma, exogenous asthma, and dusty asthma), especially chronic or refractory asthma (such as delayed asthma and tracheal overreaction), bronchitis, allergies Reversible obstructive tracheal diseases of rhinitis and the like are targets of the compounds of the invention. Inflammation of the mucosa and blood vessels includes vascular damage caused by gastric ulcers, ischemic diseases, and thrombosis. Furthermore, hyperproliferative vascular diseases including intimal smooth muscle cell proliferation, restenosis, and angiogenesis (especially following biologically or mechanically mediated vascular injury) can be treated or prevented by the compounds of the present invention.

The compounds or drugs described herein can be applied to a stent coated with a polymeric compound. The polymer coating of the compound or drug into the stent can be carried out by immersing the polymer coated stent in a solution comprising the compound or drug for a sufficient time (such as five minutes); and then (such as) The coated stent is dried by means of air drying for a sufficient time (such as 30 minutes). Other methods of applying the therapeutic substance, including spraying, can be used. The polymer coated stent comprising the compound or drug can then be delivered to the coronary vessel by deployment from a balloon catheter or via a self-expanding stent. Other devices that can be used to introduce the medicament of the present invention to the vascular structure other than the stent include, but are not limited to, grafts, catheters, and balloons. In addition, other compounds or drugs that can be used in place of the agents of the invention include, but are not limited to, A-94507 and SDZ RAD (aka Everolimus).

The compounds described herein for polymer coated stents can be used in combination with other agents. Such agents which are most effective in preventing restenosis in combination with the compounds of the present invention can be classified into anti-proliferative, anti-platelet, anti-inflammatory, antithrombotic, and thrombolytic agents. These categories can be subdivided. For example, the anti-proliferative agent can be anti-mitotic. Anti-mitotic agents inhibit or affect cell division, thereby preventing the process typically involved in cell division from occurring. One subclass of anti-mitotic agents includes vinca alkaloids. Representative examples of vinca alkaloids include, but are not limited to, vincristine, paclitaxel, etoposide, nocodazole, indirubin, and anthracycline derivatives (such as Daunol) Daunorubicin, daunomycin, and plicamycin. Other subclasses of anti-mitotic agents include anti-mitotic alkylating agents such as tauromustine, bofumustine and fotemustine, and anti-mitotic metabolites such as Amethotrexate, fluorouracil, 5-bromodeoxyuridine, 6-azacytidine nucleoside and cytarabine. Anti-mitotic alkylating agents affect cell division by covalently modifying DNA, RNA or protein thereby inhibiting DNA replication, RNA transcription, RNA translation, protein synthesis or a combination of the foregoing.

Examples of anti-mitotic agents include, but are not limited to, paclitaxel. As used herein, paclitaxel includes the alkaloid itself and its naturally occurring forms and derivatives, as well as its synthetic and semi-synthetic forms.

An antiplatelet agent is a therapeutic entity by (1) inhibiting platelet adhesion to the surface, usually the surface of prothrombin, (2) inhibiting platelet aggregation, (3) inhibiting activation of platelets, or (4) combining the foregoing effect. Activation of platelets is thereby the process of transforming platelets from a quiescent state to a state in which platelets undergo a large number of morphological changes induced by contact with the prothrombin surface. These changes include changes in platelet shape, with the formation of pseudopods (binding membrane receptors) and secretion of small molecules and proteins such as ADP and platelet factor 4. Antiplatelet agents that act as platelet adhesion inhibitors include, but are not limited to, eptifibatide, tirofiban, RGD (Arg-Gly-Asp)-based peptides that inhibit gpIIbIIIa or αvβ3 binding, Blocking antibodies that bind gpIIaIIIb or αvβ3, antibodies against P-selectin, antibodies against E-selectin, compounds that block P-selectin or E-selectin binding to their respective ligands, whey acid, and anti-Werber's (von) Willebrand) factor antibody. Agents that inhibit ADP-regulated platelet aggregation include, but are not limited to, disagregin and cilostazol.

Anti-inflammatory drugs can also be used. Examples of such anti-inflammatory agents include, but are not limited to, prednisone, dexamethasone, hydrocortisone, estradiol, triamcinolone, mometasone, Fluticasone, clobetasol and non-steroidal anti-inflammatory drugs such as acetaminophen, ibuprofen, naproxen, adalimumab and sulindac ( Sulindac). An arachidonic acid ester metabolite prostacyclin or prostacyclin analog is an example of a vasoactive anti-proliferative agent. Other examples of such agents include those that block cytokine activity or inhibit cytokines or chemokines from binding to similar receptors to inhibit pre-inflammatory signals that are converted by cytokines or chemokines. Representative examples of such agents include, but are not limited to, anti-IL1, anti-IL2, anti-IL3, anti-IL4, anti-IL8, anti-IL15, anti-IL18, anti-MCP1, anti-CCR2, anti-GM-GSF, and anti-TNF antibodies. Other examples including the second drug glucocorticol include methylprednisolone, prednisolone, prednisone, triamcinolone, dexamethasone, mometasone, beclomethasone, ciclesonide Ciclesonide, bedesonide, triamcinolone, clobetasol, flunisolide, loteprednol, budesonide, fluticasone and Salts, esters, prodrugs and derivatives thereof or any combination thereof. In other embodiments, the second drug is a steroid hormone, including estradiol and salts, esters, prodrugs and derivatives thereof, or any combination thereof.

Other agents which may be used in combination with the compounds of the invention include antilipemic agents such as fenofibrate, atorvastatin, fluvastatin, lovastatin, mevastatin (mevastatin), pitavastatin, pravastatin, rosuvastatin, and simvastatin. Other agents include matrix metalloproteinase inhibitors (such as batimistat) and endothelin A receptor antagonists (such as darusentan) and [alpha]v[beta]3 integrin receptor antagonists.

Antithrombotic agents include chemical and biological entities that can be intervened at any stage in the aggregation pathway. Examples of specific entities include, but are not limited to, small molecules that inhibit Factor Xa activity. In addition, heparin-type agents such as heparin, heparin sulfate, and low molecular weight heparin (such as having the trademark Clivarin) can be directly or indirectly inhibited. Compounds) and synthetic oligosaccharides (such as the trademark Arixtra) Compound). Also included are direct thrombin inhibitors such as melagatran, ximelagatran, argatroban, inogatran, and phenyl Phe-Pro-Arg blood of thrombin. A peptidomimetic of the binding site of the fibrinogen matrix. Another deliverable antithrombotic agent is a Factor VII/VIIa inhibitor, such as an anti-Factor VII/VIIa antibody, rNAPc2, and a tissue factor pathway inhibitor (TFPI).

A thrombolytic agent can be defined as an agent that aids in the degradation of a blood clot (clot), which can also be used as an adjuvant because the action of dissolving the clot helps to disperse the platelets that are trapped in the blood fiber matrix of the thrombus. Representative examples of thrombolytic agents include, but are not limited to, urokinase or recombinant urokinase, pro-urokinase or pre-recombinant urokinase, tissue plasminogen activator or recombinant forms thereof, and streptokinase.

Other drugs which can be used in combination with the compounds of the invention are cytotoxic drugs, such as apoptosis inducing agents (such as TGF) and topoisomerase inhibitors, including 10-hydroxycamptothecin, irinotecan and arbutin. Doxorubicin. Other types of drugs which can be used in combination with the compounds of the present invention are drugs which inhibit cell dedifferentiation and drugs which inhibit cell growth.

Other agents which may be used in combination with the compounds of the invention include antilipemic agents (such as fenofibrate), matrix metalloproteinase inhibitors (such as batimidate), antagonists of the endothelin A receptor (such as dassentan) And αvβ3 integrin receptor antagonists.

Embodiments of the invention further include a third therapeutic drug or substance. When the second drug and/or the third therapeutic drug are utilized, including but not limited to an anti-proliferative agent, an anti-platelet agent, an anti-inflammatory drug, an anti-lipid agent, an antithrombotic agent, a thrombolytic agent, a salt thereof, and a prodrug And derivatives or any combination thereof. When the second drug and/or the third therapeutic drug is a glucocorticol, it includes, but is not limited to, methylprednisolone, prednisolone, prednisone, triamcinolone, dexamethasone, mometasone, and chlorin Messon, ciclesonide, bettydine, triamcinolone, clobetasol, flunisolide, loteprednol, budesonide, fluticasone, salts, prodrugs and derivatives thereof, or any combination thereof . When the second drug and/or the third therapeutic agent is a steroid hormone, it includes, but is not limited to, estradiol and salts, prodrugs and derivatives thereof, or any combination thereof. In an embodiment, the second drug and/or the third therapeutic agent may be small molecules and biological agents that reduce the activity of inflammatory cytokines. When the second drug and/or the third therapeutic agent utilizes an anti-TNFα therapeutic agent, including, but not limited to, adalimumab, an anti-MCP-1 therapeutic agent, a CCR2 receptor antagonist, an anti-IL-18 therapeutic agent, Anti-IL-1 therapeutic agents and salts, prodrugs and derivatives thereof, or any combination thereof. When the second drug and/or the third therapeutic agent utilizes an anti-proliferative agent, it includes, but is not limited to, an alkylating agent, including cyclophosphamide, chlorambucil, busulfan Carnitustine and lomustine; antimetabolites, including methotrexate, fluorouracil, cytarabine, thioglycol and pentastatin; vinca alkaloids, including vinblastine and vinca Neobase; antibiotics, including doxorubicin, bleomycin, and mitomycin; anti-proliferative agents, including cisplatin, procarbazine, etoposide, and fentanyl Bolt; its salts, prodrugs and derivatives or any combination thereof. When the second drug and/or the third therapeutic agent utilizes an anti-platelet agent, including but not limited to glycoprotein IIB/IIIA inhibitors, including abciximab, eptifibatide, and tirofiban; gland Glycoside reuptake inhibitors, including dipyridamole; ADP inhibitors, including clopidogrel and ticlopidine; cyclooxygenase inhibitors, including acetyl sulphate; Phosphodiesterase inhibitors, including cilostazol; salts, prodrugs and derivatives thereof, or any combination thereof. When the second drug and/or the third therapeutic drug utilizes an anti-inflammatory drug, it includes, but is not limited to, steroids, including dexamethasone, hydrocortisone, fluticasone, clobetasol, mometasone, and estradiol. And non-steroidal anti-inflammatory drugs, including acetaminophen phenol, ibuprofen, naproxen, sulindac, piroxicam, mefenamic acid; inhibiting cytokines or chemokine-binding receptors Anti-inflammatory drugs that inhibit pre-inflammatory signals, including antibodies to IL-1, IL-2, IL-8, IL-15, IL-18, and TNF; salts, prodrugs, and derivatives thereof, or any combination thereof. When the second drug and/or the third therapeutic agent utilizes an antithrombotic agent, it includes, but is not limited to, heparin, including unseparated heparin and low molecular weight heparin, including clivarin, dalteparin ), enoxaparin, nadroparin, and tinzaparin; direct thrombin inhibitors, including argatroban, hirudin, hirulog, herugen Salts, prodrugs and derivatives thereof or any combination thereof. When the second drug and/or the third therapeutic agent utilizes an antilipemic agent, it includes an HMG CoA reductase inhibitor, including mevastatin, lovastatin, simvastatin, pravastatin, fluvastatin; , including fenofibrate, clofibrate, gemfibrozil; lipid lowering agents, including nicotinic acid, probucol; salts, prodrugs and derivatives thereof, or any combination thereof . When the second drug and/or the third therapeutic drug utilizes a thrombolytic agent, it includes, but is not limited to, a hammer stimulation, urokinase, prourokinase, tissue plasminogen activator (including alteplase) , reteplase, tenectaplase, salts, prodrugs and derivatives thereof, or any combination thereof.

polymer

When used in the present invention, the coating can comprise any polymeric material in which the therapeutic agent (i.e., drug) is substantially soluble or effective. The purpose of the coating is to act as a controlled release vehicle for the therapeutic agent or as a reservoir for the therapeutic agent delivered at the site of injury. The coating can be polymeric and can be further hydrophilic, hydrophobic, biodegradable or biodegradable. The material used for the polymeric coating may be selected from the group consisting of polycarboxylic acids, cellulosic polymers, gelatin, polyvinylpyrrolidone, maleic anhydride polymers, polyamines, polyvinyl alcohol, Polyethylene oxide, glycosaminoglycan, polysaccharide, polyester, polyurethane, polyoxo, polyorthoester, polyanhydride, polycarbonate, polypropylene, polylactic acid, polyglycolic acid, polyhexyl A mixture and copolymer of a lactone, a polyhydroxybutyrate valerate, a polypropylene decylamine, a polyether, and the foregoing polymers. Coatings prepared from polymeric dispersions, including polyurethane dispersions (BAYHYDROL, etc.) and acrylic latex dispersions, may also be used with the therapeutic agents of the present invention.

Biodegradable polymers useful in the present invention include polymers including poly(L-lactic acid), poly(DL-lactic acid), polycaprolactone, poly(hydroxybutyrate), polyacetate. Ester, dixanone, poly(hydroxyvalerate), polyorthoester, copolymer (including poly(lactide-co-glycolide), polyhydroxy (butyrate-co-valerate) , polyglycolide-co-trimethylene carbonate), polyanhydrides, polyphosphates, polyphosphates-urethanes, polyamino acids, polycyanoacrylates, biomolecules (including blood fibers, A mixture of fibrinogen, cellulose, starch, collagen, and hyaluronic acid) and the foregoing polymers. Biostable materials suitable for use in the present invention include polymers comprising the following polymers: polyurethane, polyoxo, polyester, polyolefin, polyamine, polycaprolactam, polyimine, poly Vinyl chloride, polyvinyl methyl ether, polyvinyl alcohol, acrylic polymer and copolymer, polyacrylonitrile, polystyrene copolymer of olefinic monomer and olefin (including styrene acrylonitrile copolymer, methacrylic acid Methyl ester copolymer, ethyl vinyl acetate), polyether, human cotton, cellulose (including cellulose acetate, nitrocellulose, cellulose propionate, etc.), parylene and its derivatives and Mixtures and copolymers of the foregoing polymers.

(Steve provides a variety of genetically included Guidant polymers. In some embodiments, the polymer includes, but is not limited to, poly(acrylate) (such as poly(ethyl methacrylate), poly(methacrylic acid) N-propyl ester), poly(isopropyl methacrylate), poly(isobutyl methacrylate), poly(butyl methacrylate), poly(n-butyl methacrylate), poly(methyl 2-ethylhexyl acrylate), poly(n-hexyl methacrylate), poly(cyclohexyl methacrylate), poly(n-hexyl methacrylate), poly(isobornyl methacrylate) and poly( Trimethylcyclohexyl methacrylate), poly(methyl acrylate), poly(ethyl acrylate), poly(n-propyl acrylate), poly(isopropyl acrylate), poly(n-butyl acrylate), Poly(isobutyl acrylate), poly(second butyl acrylate), poly(pentyl acrylate), poly(n-hexyl acrylate), poly(cyclohexyl acrylate), and any derivatives, analogs, homologs, Homologues, salts, copolymers, and combinations thereof.

In some embodiments, the polymer includes, but is not limited to, poly(ester urethane), poly(ether urethane), poly(ureidourethane), poly(amino carboxylic acid) Triester copolymer of styrene and isobutylene, copolymers of polyoxymethylene, fluoropolyoxygen, poly(ester), poly(ethylene), poly(propylene), poly(olefin), poly(isobutylene) , a triblock copolymer of styrene and ethylene/butylene, a triblock copolymer of styrene and butadiene, a copolymer of ethylene-alpha olefin, a halogenated ethylene polymer and a copolymer (such as poly(ethylene chloride) And poly(ethylene fluoride)), poly(halogenated vinylene) (such as poly(vinylidene) and poly(fluoroethylene)), poly(vinylidene fluoride-co-hexafluoropropylene), poly(tetrafluoroethylene) , poly(tetrafluoroethylene-co-chlorotrifluoroethylene), poly(vinyl ether) (such as poly(vinyl methyl ether)), poly(acrylonitrile), poly(vinyl ketone), poly (vinyl aromatic) a compound (such as poly(styrene)), a poly(vinyl ester) (such as poly(vinyl acetate)), a copolymer of an olefinic monomer and an olefin (such as a copolymer of methacrylic acid, a copolymer of acrylic acid) , a copolymer of N-vinylpyrrolidone, Poly(vinyl alcohol), poly(ethylene-co-vinyl alcohol) (EVAL), poly(cyanoacrylate), copolymer of poly(maleic anhydride) and maleic anhydride, acrylonitrile- Copolymer of styrene, copolymer of ABS resin and ethylene-vinyl acetate) and any derivatives, analogs, homologs, homologs, salts, copolymers and combinations thereof.

In some embodiments, the polymer includes, but is not limited to, poly(decylamine) (such as Nylon 66 and poly(caprolactam)), alkyd, poly(carbonate), poly(碸) ), poly(formaldehyde), poly(indenine), poly(esteramine), poly(ether) (including poly(alkylene glycol) such as poly(ethylene glycol) and poly(propylene glycol), epoxy Resin, human cotton, human cotton-triacetate, biomolecules (such as blood fiber, fibrinogen, starch, poly(amino acid), peptide, protein, gelatin, chondroitin sulfate, dermatan sulfate (D- Glucuronic acid or a copolymer of L-iduronic acid with N-ethinyl-D-galactosamine), collagen, hyaluronic acid and glycosaminoglycan), other glycans (such as poly(N) -Ethyl glucosamine), chitin, polyglucosamine, cellulose, cellulose acetate, cellulose butyrate, cellulose acetate butyrate, cellophane, nitrocellulose, cellulose propionate , cellulose ethers and carboxymethyl cellulose) and any derivatives, analogs, homologs, homologs, salts, copolymers and combinations thereof.

Another polymer useful in the present invention is poly(MPC _w : LAM _x : HPMA _y : TSMA _z ), where w, x, y, and z represent the molar ratio of the monomers used in the feedstock used to prepare the polymer. And MPC represents the unit 2-methylpropenyloxyethylphosphonium choline, LMA represents the unit of lauryl methacrylate, HPMA represents the unit 2-hydroxypropyl methacrylate, and TSMA represents the unit methacrylic acid 3-trimethoxydecylpropyl propyl ester. Drug-impregnated stents can be used to maintain the opening of coronary arteries previously occluded by thrombus and/or atherosclerotic plaques. Delivery of anti-proliferative agents reduces the rate of restenosis under the stent. Polymers useful in the present invention include zwitterionic polymers including phosphonium choline units.

Other treatable conditions include, but are not limited to, ischemic enteropathy, inflammatory bowel disease, necrotizing enterocolitis, enteritis/allergy (including celiac disease, proctitis, eosinophilic gastroenteritis, mastocytosis, Crohn's disease and ulcerative colitis), neuropathy (including polymyositis, Guillain-Barre syndrome, Meniere's disease, polyneuritis, multiple nerves) Inflammation, mononeuritis and radiculopathy), endocrine diseases (including hyperthyroidism and Basedow disease), blood diseases (including pure red blood cell aplastic anemia, aplastic anemia, low-forming anemia, Idiopathic thrombocytopenic purpura, autoimmune hemolytic anemia, granulocytopenia, pernicious anemia, macroglobulin anemia, and red blood cell insufficiency), bone disease (including osteoporosis), respiratory disease (including sarcoidosis) , fibrous lung and idiopathic interstitial pneumonia), skin diseases (including dermatomyositis, leukoplakia, common ichthyosis, photoallergic sensitivity and cutaneous T-cell lymphoma), circulatory diseases (including arteries) Syndrome, atherosclerosis, aortic inflammatory syndrome, nodular polyarteritis and non-inflammatory cardiomyopathy), collagen disease (including scleroderma, Wegener granulomatosis and Sjogren syndrome) ), obesity, eosinophilic fasciitis, periodontal disease (including gum, root canal, alveolar bone and cementum damage), renal syndrome (including spheroid nephritis), male alopecia or alopecia premature aging ( By preventing hair loss or providing hair growth and/or promoting hair growth and hair growth), muscular dystrophy, pyoderma and Sezary syndrome, Addison disease, ROS-regulating diseases (eg organs Injury, including organ (including heart, liver, kidney and digestive tract) ischemia-reperfusion injury (which occurs during protection, transplantation) or ischemic diseases (such as thrombosis and myocardial infarction), intestinal diseases (including Toxin-shock, pseudomembranous colitis and colitis caused by drugs or radiation), kidney disease (including ischemic acute renal insufficiency and chronic renal insufficiency), lung disease (including lung-oxygen or drugs (Pacocote ( Toxin disease caused by paracort) and bleomycin Carcinoma and emphysema), eye diseases (including cataract, calculus, retinitis pigmentosa, age-related macular degeneration, vitreous crust and corneal alkali burn), dermatitis (including polymorphous erythema, linear IgA sclerotia and Cement dermatitis) and others, including gingivitis, periodontitis, sepsis, pancreatitis, diseases caused by environmental pollution (such as air pollution), old age, carcinogenesis, cancer metastasis, and hypobaric disease, Disease caused by histamine or leukotriene-C ₄ release, Behcet's disease (including intestinal, vascular or neurobehide disease and white affecting the mouth, skin, eyes, vulva, joints, epididymis, lungs, kidneys, etc.) Say's disease). In addition, the compounds of the present invention are useful for the treatment and prevention of liver diseases, including immunogenic diseases (such as chronic autoimmune liver disease, including autoimmune hepatitis, primary biliary cirrhosis, and sclerosing cholangitis), partial hepatectomy. Acute hepatic necrosis (eg necrosis caused by toxins, viral hepatitis, shock or anoxia), B-viral hepatitis, non-A/non-B hepatitis, sclerosis (including alcoholic cirrhosis) and liver failure ( These include fulminant hepatic failure, delayed onset liver failure, and "chronic acute" liver failure (acute liver failure on chronic liver disease), and in addition to the major chemical effects, antiviral effects of these compounds on the drugs that patients may use, The anti-inflammatory effect and the expansion of the cardiotonic effect have potentially useful activities and are therefore suitable for a variety of diseases.

The ability of the compounds of the invention to treat proliferative diseases can be as previously described in Bunchman ET and CA Brookshire, Transplantation Proceed. 23 967-968 (1991); Yamagishi et al, Biochem. Biophys. Res. Comm. 191 840-846 (1993); This is demonstrated by the method described in Shichiri et al., J. Clin. Invest. 87 1867-1871 (1991). Proliferative diseases include smooth muscle hyperplasia, systemic sclerosis, cirrhosis, adult respiratory distress syndrome, idiopathic cardiomyopathy, lupus erythematosus, diabetic retinopathy or other retinopathy, psoriasis, scleroderma, benign prostatic hyperplasia, cardiac hyperplasia, Restenosis after arterial injury or other pathological stenosis of blood vessels. In addition, these compounds antagonize cellular responses to some growth factors and thus possess anti-angiogenic properties, which makes them useful agents for controlling or reversing certain tumor growth and fibrotic diseases of the lung, liver and kidney.

The aqueous liquid compositions of the embodiments of the present invention are particularly useful for the treatment and prevention of various ocular diseases, including autoimmune diseases including, for example, keratoconus, keratitis, corneal epithelial dystrophy, corneal leukoplakia, Mooren ulcers. , sclevitis and Graves' eye lesions) and corneal transplant rejection.

When used in the above or other treatments, a therapeutically effective amount of one of the compounds of the embodiments of the invention may be used neat or in the form of a pharmaceutically acceptable salt, ester or prodrug if present. . Alternatively, the compound can be administered as a pharmaceutical composition comprising a combination of the relevant compound and one or more pharmaceutically acceptable excipients. The phrase "therapeutically effective amount" of a compound of the invention means a compound that is sufficient to treat a condition in a reasonable benefit/risk ratio applicable to any medical treatment. However, it is to be understood that the total daily usage of the compounds and compositions of the present invention is determined by the attending physician within the scope of sound medical judgment. The particular therapeutically effective dose for any particular patient will depend on a number of factors, including the severity of the condition and condition being treated, the activity of the particular compound employed, the particular composition employed, the patient's age, weight, and overall Health, sex and diet, time of administration, route of administration and rate of excretion of the particular compound used, duration of treatment, combination or consistency with the particular compound employed, and similar factors well known in the medical arts. For example, it is entirely within the skill to start the dosage of the compound in an amount lower than that required to achieve the desired therapeutic effect and gradually increase the dosage until the desired effect is achieved.

The total daily dose of a compound of the present invention administered to a human or lower animal can vary from about 0.01 to about 10 mg/kg per day. For the purpose of oral administration, the dosage may range from about 0.001 to about 3 mg/kg per day. For the purpose of local delivery from the stent, the daily dose received by the patient depends on the length of the stent. For example, a 15 mm coronary stent can include a drug that varies from about 1 to about 600 mg, and can deliver the drug over a period of time ranging from several hours to several weeks. If desired, the effective daily dose can be divided into multiple doses for the purpose of administration, and thus, a single dose of the composition can include such amounts or submultiples thereof to form a daily dose. For topical administration, the present invention can be used by those skilled in the art and the dosage will depend on the site of application.

Within the scope of the present invention, a suitable polymer layer for loading a drug is provided with great flexibility. For example, within the therapeutic range parameters associated with the compound of interest (typically between therapeutic efficacy and toxicity), the ratio of the compounds used in combination can vary relative to each other. For example, an embodiment has a total drug:polymer ratio of 90:10, wherein the ratio of drugs in the combination can be 1:1. Thus, the stent of the present invention for delivering a quetiamus/dexamethasone combination can comprise 10 mcg/mm zetamus and 10 mcg/mm in a PC polymer layer having a 5 mcg/mm PC overcoat. Dexamethasone. However, the total drug:polymer ratio can be lower, such as 40:60 or less. The upper limit of the total amount of the drug depends on a number of factors, including the miscibility of the selected drug in the selected polymer, the stability of the drug/polymer mixture (eg, bactericidal compatibility), and the physical properties of the mixture. (eg fluidity/handleability, elasticity, brittleness, viscosity (no mesh or bridge formed between the stent rods), coating thickness (substantially increasing the stent profile or causing delamination or cracking or difficulty curling). Embodiments of the invention include stent rods spaced about 60-80 microns apart, which indicates that the drug/polymer/polymer overcoat has an upper thickness limit of about 30 microns; however, for drug delivery as described elsewhere, Use any bracket size, rod size and spacing and/or bracket structure. In an embodiment, the therapeutic amount of the Moss drug comprises quetiamus or everolimus, and is at least 1 μg per millimeter of stent. In other embodiments, the second drug is a glucocorticol. When the second drug is utilized in the embodiment, the second drug is dexamethasone and the therapeutic amount is at least 0.5 μg per millimeter of stent.

It is expected that the thickness of the outer coating (if an outer coating is used) should not unduly impede the release kinetics of the drug. The overcoat layer may also be loaded with one or more drugs which may be the same or different from the drug in the polymer layer supporting the base drug.

In general, combining the drugs used in the present invention does not adversely affect the desired activity of other drugs in the combination. The proposed drug for use in the combination may have a complementary activity or mechanism of action. Thus, one of the proposed combinations should not inhibit the desired activity, such as the anti-proliferative activity of another drug. No drug should cause or enhance the degradation of another drug. However, due to, for example, degradation during sterilization, a seemingly unsuitable drug may actually be useful due to the interaction of another drug. Therefore, dexamethasone which was degraded during EtO sterilization when used alone was observed to be successfully used in combination with quetiamus because of the hydrophobicity of quetiamus. In addition, it has been observed that quetiamus reduces the rate of dissolution of dexamethasone, as described in U.S. Patent Application Serial No. 10/796,423, the entire disclosure of which is incorporated herein by reference.

The pharmaceutical composition of the embodiments of the present invention comprises a compound and a pharmaceutically acceptable carrier or excipient which can be orally, rectally, parenterally, cerebrally, intravaginally, intraperitoneally, locally (e.g. Administration or delivery to the pericardial space by powder, ointment, drip or dermal patch, buccal (eg oral or nasal spray) or topical (eg in a stent placed in a vascular structure such as in a balloon catheter) Transfer to or within the myocardium. The phrase "pharmaceutically acceptable carrier" means a non-toxic solid, semi-solid or liquid filler, diluent, sealing material or formulation aid of any type. The term "parenteral" as used herein refers to all modes of administration other than oral administration including, but not limited to, intravenous, intraarterial, intramuscular, intraperitoneal, intrasternal, subcutaneous and intraarticular injection, drip, Percutaneously and placed (such as within a vascular structure).

The pharmaceutical composition of the present invention for parenteral injection comprises a pharmaceutically acceptable sterile aqueous or nonaqueous solution, dispersion, suspension, nanoparticle suspension or emulsion and reconstituted as a sterile injectable solution or reconstituted before use or A sterile powder of the dispersion. Examples of suitable aqueous and non-aqueous vehicles, diluents, solvents or vehicles include water, ethanol, polyols (including glycerol, propylene glycol, polyethylene glycol, and the like), carboxymethylcellulose, and suitable mixtures thereof, Vegetable oils (including olive oil) and injectable organic esters (including ethyl oleate). Suitable fluidity can be maintained, for example, by the use of a coating material comprising lecithin, by the maintenance of the desired particle size under the conditions of the dispersion, and by the use of a surfactant.

Such compositions may also include adjuvants such as preservatives, wetting agents, emulsifying agents and dispersing agents. Prevention of the action of microorganisms can be ensured by including various antibacterial agents and antifungal agents such as parabens, chlorobutanol, phenol sorbic acid and the like. It is also desirable to include isotonic agents, including sugars, sodium chloride, and the like. Prolonged absorption of the pharmaceutical form of the injectables can be brought about by the inclusion of agents which comprise extended absorption, including aluminum monostearate and gelatin.

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 a liquid suspension of crystalline or amorphous material having poor water solubility. Subsequently, the rate of absorption of the drug depends on its rate of dissolution, which in turn depends on the crystal size and crystalline form. Alternatively, delayed absorption of a parenterally administered drug form is accomplished by dissolving or suspending the drug in an oil vehicle.

The injectable reservoir form is made by forming a microcapsule matrix of the drug in a biodegradable polymer, including polylactide-polyglycolide, depending on the ratio of drug to polymer and the nature of the particular polymer used. The drug release rate can be controlled. Examples of other biodegradable polymers include poly(orthoesters) and poly(anhydrides). The reservoir injection formulation can also be prepared by incorporating the drug into liposomes or microemulsions that are compatible with body tissues.

The injectable formulation can be sterilized, for example, by passing through a filter that blocks the bacteria or by incorporating the bactericide into a sterile solid composition form that is soluble or dispersible in sterile water just prior to use. Or other sterile injectable media.

Solid dosage forms for oral administration include capsules, lozenges, pills, powders and granules. In such solid dosage forms, the active compound is combined with at least one inert, pharmaceutically acceptable excipient or carrier (including sodium citrate or dicalcium phosphate) and/or a) filler or excipient (including starch, Lactose, sucrose, glucose, mannitol and citric acid), (b) binders (such as carboxymethylcellulose, alginates, gelatin, polyvinylpyrrolidone, sucrose and gum arabic), c) humectants (including Glycerin), d) disintegrants (including agar, calcium carbonate, potato or tapioca starch, alginic acid, certain citrates and sodium carbonate), e) solution retarders (including paraffin), f) absorption enhancers (including a quaternary amine compound), g) a wetting agent (such as cetyl alcohol and glyceryl monostearate), h) an adsorbent (including kaolin and bentonite), and i) a lubricant (including talc, calcium stearate, hard) Magnesium citrate, solid polyethylene glycol, sodium lauryl sulfate, and mixtures thereof are mixed. In the case of capsules, lozenges and pills, the dosage form may also contain a buffer.

Solid compositions of a similar type may also be employed as fillers in soft, semi-solid and hard-filled gelatin capsules or in liquid-filled capsules (using excipients such as lactose and high molecular weight polyethylene glycols and the like) Agent.

Solid dosage forms (not limited to lozenges, dragees, capsules, pills, and granules) having a coating and shell comprising other coatings well known in the casing and pharmaceutical formulation techniques can be prepared for oral administration. It may optionally contain a creaming agent and may also be a composition that only (or preferentially) releases the active ingredient in an extended manner, as appropriate, in a particular portion of the intestinal tract. Examples of embedding compositions that can be used include polymeric materials and waxes. The embedding compositions comprising the drug can be placed on a medical device comprising a stent, a graft, a catheter and a balloon.

The active composition may also be in the form of microcapsules, if appropriate, having one or more of the excipients described above.

Liquid dosage forms for oral administration include pharmaceutically acceptable emulsions, solutions, suspensions, syrups and spirits. Liquid dosage forms can include, in addition to the active compound, inert diluents commonly employed in the art, such as water or other solvents, solubilizers and emulsifiers, including ethyl alcohol, isopropyl alcohol, ethyl carbonate, ethyl acetate, Benzyl alcohol, benzyl benzoate, propylene glycol, 1,3-butanediol, dimethylformamide, oil (especially cottonseed oil, peanut oil, corn oil, germ oil, olive oil, castor oil and sesame oil), glycerin , fatty acid esters of tetrahydrofurfuryl alcohol, polyethylene glycol and sorbitan, and mixtures thereof.

Besides inert diluents, the oral compositions can also include adjuvants including humectants, emulsifying and suspending agents, sweetening agents, flavoring agents, and flavoring agents.

In addition to the active compound, the suspension may include suspending agents such as ethoxylated isostearyl alcohol, polyoxyethylene sorbitol and sorbitan esters, microcrystalline cellulose, aluminum metahydroxide, bentonite, Agar and tragacanth and their mixtures.

Topical administration includes administration to the skin including the surface of the eye. Compositions for topical application to the skin also include ointments, creams, lotions and gels. Another form of topical administration is administration to the eye, such as immunomodulatory conditions for the treatment of the eye (including autoimmune diseases, allergic or inflammatory conditions, and corneal transplants). The compounds of the invention are delivered in a pharmaceutically acceptable ophthalmic vehicle such that the compound remains in contact with the surface of the eye for a time sufficient for the compound to penetrate the cornea and internal regions of the eye, such as the anterior chamber, posterior chamber, vitreous, water Fluid, vitreous, cornea, iris / ciliary body, lens, choroid / retina and sclera. Pharmaceutically acceptable ophthalmic vehicles can, for example, be ointments, vegetable oils or encapsulated materials.

Compositions for mucosal administration, especially for inhalation, can be prepared as dry powders which may or may not be pressurized. In a non-pressurized powder composition, the active ingredient in the form of a fine powder may be mixed with a larger pharmaceutically acceptable inert carrier (including particles having a size of, for example, up to 100 microns in diameter) use. Suitable inert carriers include sugars, including lactose. It is desirable that at least 95% by weight of the active ingredient particles have an effective particle size in the range of from 0.01 to 10 microns. In transrectal or vaginal administration via the mucosa, the formulation includes a suppository or retention enemas which can be prepared by combining a compound of the invention with a suitable non-irritating excipient comprising cocoa butter, polyethylene glycol or suppository wax or The agent, which is solid at room temperature but liquid at body temperature and thus melts in the rectum or vaginal cavity and releases the active compound, is prepared by mixing.

Alternatively, the composition is pressurized and includes a compressed gas, including nitrogen or a liquefied gas propellant. The liquefied propellant medium and in fact the total composition is a liquefied propellant medium and in fact a total composition in which the active ingredient is not dissolved in any substantial extent. The pressurized composition may also include a surfactant. The surfactant can be a liquid or solid nonionic surfactant or can be a solid anionic surfactant. In other embodiments, the use of a solid anionic surfactant is in the form of a sodium salt.

The compounds of the examples of the invention may also be administered in the form of liposomes. As is known in the art, liposomes are typically obtained from phospholipids or other lipid materials. Liposomes are formed by hydrating liquid crystals of single or multiple flakes dispersed in an aqueous medium. Any non-toxic, physiologically acceptable and metabolizable lipid capable of forming liposomes can be used. In addition to the compounds of the present invention, examples of compositions in liposome form may include stabilizers, preservatives, excipients, and the like. The lipids in the examples are natural and synthetic phospholipids and phospholipids choline (lecithin). Methods of forming liposomes are known in the art. See, for example, the book Prescott, Methods in Cell Biology. Volume XIV, Academic Press, New York, NY (1976), page 33 and the following.

The compounds of the embodiments of the invention may also be administered in combination with one or more systemic immunosuppressive agents. Immunosuppressive agents within the scope of the invention include, but are not limited to, IMURAN Azathioprine sodium, brequinar sodium, SPANIDIN Gusperimus trihydrochloride (also known as deoxyspergualin), imidazole (mizoribine) (also known as bredinin), CELLCEPT Mycophenolate, NEORAL Cyclosporin A (also in the trademark SANDIMMUNE Different formulations of cyclosporin A purchased under), PROGRAF Tacrolimus (also known as FK-506), sirolimus and RAPAMUNE Everolimus, leflunomide (also known as HWA-486), glucocorticoids (including splashed nylon and its derivatives), antibody therapeutics (including orthoclone (OKT3)), and Zenapax , leukemia therapeutic agents and anti-thymocyte globulin (including rabbit thymoglobulin).

Preparation of the compounds of the invention

The compounds and methods of the examples of the present invention are better understood in conjunction with the following synthetic schemes for the preparation of the compounds of the present invention.

The compounds of the invention can be prepared by a variety of synthetic routes. A representative method is shown in Scheme 1.

As shown in Scheme 1, the C-42 hydroxyl group of rapamycin is converted to a triflate or fluorosulfonate leaving group to give A. Substituting the leaving group with a tetrazole in the presence of a non-nucleophilic hindered base (including 2,6-lutidine, diisopropylethylamine) to give isomers B and C, isomer B and C It was separated and purified by flash column chromatography.

resolve resolution

The foregoing is to be understood by reference to the description of the accompanying claims

Example 1 42-(2-tetrazolyl)-rapamycin (the less polar isomer)

Example 1A A solution of rapamycin (100 mg, 0.11 mmol) in dichloromethane (0.6 mL) was used in a solution of 2,6-dimethylpyridine (53 μL, 0.46 mmol) at -78 ° C under nitrogen atmosphere. Treated with trifluoromethanesulfonic acid anhydride (37 μL, 0.22 mmol), and then stirred for 15 min, warmed to room temperature and eluted with a pad of diethyl ether (6 mL). The fractions including the triflate are concentrated and concentrated to yield the designated compound as an amber foam.

Example 1B 42-(2-Tetrazolyl)-rapamycin (a less polar isomer) A solution of Example 1A in isopropyl acetate (0.3 mL) was sequentially taken from diisopropylethylamine (87) Treatment with mL, 0.5 mmol) and 1H-tetrazole (35 mg, 0.5 mmol), and then stirred for 18 h. This mixture was partitioned between water (10 mL) and ether (10 mL). The organics were washed with brine (10 mL) and dry (Na ₂ SO ₄₎ and washed. Concentrated organics gave a viscous yellow solid by using hexanes (10 mL), hexanes: ether (4:1 (10 mL), 3:1 (10 mL), 2:1 (10 mL), 1: It was purified by chromatography on 1 (10 mL)), ether (30 mL), hexane: acetone (1:1 (30 mL)). One of the isomers is collected in the ether fraction.

MS (ESI) m / e 966 (M) ^-

Example 2 42-(1-Tetrazolyl)-rapamycin (more polar isomer)

The slower moving band of the column was collected from the hexane:acetone (1:1) mobile phase from Example 1B to yield the indicated compound.

MS (ESI) m / e 966 (M) ^-

Pharmacokinetics of rapamycin analogues Immunosuppressive activity of compounds of the examples of the invention with rapamycin and two rapamycin analogues: 40-epi-N-[2'-pyridone]- Rapamycin and 40-epi-N-[4'-pyridone J-rapamycin, both of which are disclosed in U.S. Patent No. 5,527,907. The activity was determined using the Human Mixed Lymphocyte Reaction (MLR) assay described by Kino, T. et al., Transplantation Proceedings, XIX (5): 36-39, 6th Supplement (1987). As shown in Table 1 , the assay results demonstrate that the compounds of the invention are effective immunomodulators at the concentration of the nanomolar.

The pharmacokinetic behavior of Example 1 and Example 2 was characterized according to a single 2.5 mg/kg intravenous dose in the stone crab macaque (n=3 per group). Each compound was prepared as 20% ethanol: 30% propylene glycol: 2% cetyl polyoxyethylene ether EL: 48% dextrose 5% solution in water vehicle 2.5 mg/mL. A 1 mL/kg intravenous dose was injected slowly (about 1-2 minutes) into the saphenous vein of the monkey. Blood samples were obtained from the femoral artery or vein of each animal prior to dosing and at 0.1 (intravenous), 0.25, 0.5, 1, 1.5, 2, 4, 6, 9, 12, 24, and 30 hours after dosing. The EDTA protected samples were thoroughly mixed and extracted for subsequent analysis.

An aliquot of blood (1.0 mL) was hemolyzed with 20% methanol (0.5 mL) in water containing the internal standard. The dissolved sample was extracted with a mixture of ethyl acetate and hexane (1:1 (v/v), 6.0 mL). The organic layer was evaporated to dryness with a stream of nitrogen at room temperature. The sample was rehydrated in methanol: water (1:1, 150 μL). The title compound (50 μL injection) was separated from the contaminant by reverse phase HPLC with UV detection. The sample was kept at low temperature (4 ° C) throughout the process. All samples from each study were analyzed as a single batch on HPLC.

The area under the curve (AUC) of Example 1, Example 2, and the internal standard was determined using the Sciex MacQuan ^{T M} software. A calibration curve was obtained from the peak area ratio (parent drug/internal standard) of the cone blood standard using the least squares linear regression of the relationship between the ratio and the theoretical concentration. Within the range of the standard curve (these methods are linear for both compounds with a correlation greater than 0.99), the estimated quantitative limit is 0.1 ng/mL. The maximum blood concentration (C _{M A X} ) and the time to maximum blood concentration (T _{M A X} ) were read directly from the observed blood concentration-time data. Blood concentration data was submitted to a multi-exponential curve device using CSTRIP to obtain an assessment of pharmacokinetic parameters. The parameters evaluated are further defined using NONLIN84. The area under the blood concentration-time curve (AUC _{0 - t} ) from 0 to t hours (the last time point at which blood concentration can be measured) after administration was calculated using a linear trapezoidal rule for the blood-time curve. The remaining area extrapolated to infinity is determined as the final measured blood concentration (C _t ) divided by the terminal elimination rate constant (β), and AUC _{0 - t is added} to produce the total area under the curve (AUC _{0 - t} ).

Shorter terminal as shown in FIG. 1 and Table 2, when compared to rapamycin, Example 1 and Example 2 have substantially the surprising elimination half-life (t _1/2). Thus, only the compounds of the invention provided sufficient efficiency (Table 1) and shorter elimination half-lives ( Table 2 ).

Example 3

The purpose of this example was to determine the effect of rapamycin analogues on neovascular intimal formation in porcine coronary arteries including stents. This example demonstrates that when the rapamycin analogue quetiamus is mixed and delivered from a biocompatible BiodiviYsio PC coronary stent, it beneficially affects neovascular intimal hyperplasia and lumen size in the porcine coronary arteries. This finding suggests that if properly applied to humans, the delivery of quetiamus from a medical device is substantially clinically beneficial by limiting neovascular intimal hyperplasia.

The agent ceramimus is a rapamycin analogue, which is described and claimed in U.S. Patent No. 6,015,815. The studies presented in this example were constructed to assess the ability of the rapamycin analogue, quetiamus, to reduce neointimal hyperplasia in a porcine coronary stent model. The efficiency of quetiamus in this model indicates that quetiamus limits the clinical potential of coronary and vascular restenosis in the stent after percutaneous revascularization. The use of domestic pigs is due to the fact that this model appears to be comparable to other studies that seek to limit neovascular intimal hyperplasia in human subjects.

The examples were tested for the dissolution of the coronary stent from the juvenile farm pigs and compared to the control stents. The control scaffold is a drug-free coated polymer. Importantly, the polymer itself does not substantially stimulate neovascular intimal hyperplasia. Since the dissolving drug disappears, the inflammatory response to the polymer can imaginably lead to a late "catch up phenomenon" in which the restenosis process is not stopped, but is subdued. This phenomenon leads to restenosis in the data of human subjects.

The stent was implanted into two of the blood vessels of each pig. Pigs used in this model are typically 2-4 months old and weigh 30-40 Kg. Therefore, two coronary stents were implanted into each pig by visually evaluating the usual stent: arterial ratio at 1.1-1.2. Beginning on the day of the start of the procedure, pigs were given aspirin (325 mg daily) and continued for the remainder of the procedure. General anesthesia was achieved by intramuscular injection followed by intravenous ketamine (30 mg/kg) and xylazine (3 mg/kg). At the time of induction, intramuscular administration included additional drugs such as atropine (1 mg) and flosilillin (1 g). During the stenting procedure, a rapid injection of 10,000 units of heparin was administered.

The artery is accessed by reducing the right external carotid artery and placing an 8F sheath. After the procedure, the animals are raised on a normal diet without cholesterol or other special supplements.

A BiodivYsio stent with a target size of 3.0 mm was used. See Figure 2 . The two coronary arteries of each pig were randomly assigned to deploy the stent. The stent is a drug-dissolving stent (polymer plus drug stent) or a polymer-coated stent (polymer-only stent). The stent is delivered by means of a standard guide tube and wire. The stent balloon is inflated to a suitable size for less than 30 seconds.

Each pig has a polymer-only scaffold and a polymer plus drug scaffold placed in the isolated coronary artery such that each pig has a drug scaffold and a control scaffold.

Sample sizes of 20 total pigs were selected to detect salient differences in neovascular intimal thickness at a power of 0.95 and 0.12 mm at β 0.05 with a standard deviation of 0.15 mm.

Animals were euthanized for 28 days for histopathological examination and quantification. After the heart is removed from the perfusion pump system, the left atrial appendage is removed to access the proximal coronary artery. Dissect the coronary artery segment with injury so that there is no epicardium. The segment containing the lesion is separated whereby sufficient tissue is included at each end to include uncontained blood vessels. The aforementioned sections each having a length of 2.5 cm were embedded and treated by means of standard plastic embedding techniques. Tissues were then processed and stained with hematoxylin-eosin and elastic-van Gieson techniques.

The length measurements were performed on the plane of the micrograph using low and high power optical microscopy using a digital microscopy system with calibration calibration lines and a computer connected to the computer using the calibrated analysis software.

The severity of vascular injury and neovascular intimal response were measured by calibrated digital microscopy. Those skilled in the art are familiar with the importance of the integrity of the inner elastic layer. The histopathological damage score in the vessel with the stent has been confirmed due to its close correlation with neovascular intima thickness. This score is related to the depth of damage and is as follows:

Quantitative measurements of lesions were evaluated for all stent rods of each stent section. Neovascular intima thickness was also measured at each stent rod point using a calibrated digital image. The area of the inner cavity, the area under the inner elastic thin layer and the area within the outer elastic thin layer are also measured.

The middle bracket section is used for measurement, analysis and comparison. Information about the proximal and distal segments is also recorded (and is included in the information section of this report).

Paired t-tests were performed to compare variables across the polymer-only scaffold (control) and the polymer plus drug scaffold (treatment group). In this study, no animal died before the scheduled time.

Table 3 shows the pigs and arteries used. In Table 3 , LCX means the circumflex artery of the left coronary artery, LAD means the left anterior descending coronary artery, and RCA means the right coronary artery.

Table 4 shows an overview of all the data on the mean lesion and neovascular intima thickness of each stent (including the proximal, middle, and distal segments). Table 4 also shows lumen dimensions, percent stenosis, and arterial size as measured by the inner elastic thin layer (IEL) and the outer elastic thin layer (EEL).

There were no statistically significant differences in neovascular intimal area or thickness between the test group (polymer plus drug stent) or the control group (polymer-only stent) in the proximal, middle or distal segments. This observation is very consistent with previous studies, thus allowing a statistical comparison of the test device (polymer plus drug scaffold) to the control device (polymer only scaffold) using only the middle segment.

Table 5 shows a statistical t-test comparison across the test and control groups. There were statistically significant differences in neovascular intimal thickness, neovascular vascular area, luminal size, and percentage of luminal stenosis. Drug-dissociated stents were significantly beneficial. In contrast, there was no statistically significant difference between the test group (polymer plus drug stent) and the control group (polymer only stent) with respect to the mean damage fraction, the outer elastic thin layer or the inner elastic thin layer area. The difference.

The reference arteries on the proximal and distal sides of the stretched segment are observed and quantified. These vessels appeared normal in all conditions and were not damaged in the control group (polymer stent only) and the test group (polymer plus drug stent). See Figures 3A and 3B . The following data shows that there is no statistically significant difference in size between the stent in the control group and the stent in the test group.

The data proves that there is a statistically significant difference in the morphometric measurement of efficiency, preferring the stent of the solute. The stent of the present invention results in a lower neovascular intimal area, a lower neovascular intima thickness, and a larger lumen area. There were no significant differences in inflammatory or insult parameters between the test group (polymer plus drug stent) and the control group (polymer only stent). There was no significant difference in arterial size (including stents) compared to the test group. These recent findings indicate that there are no significant differences in the arterial remodeling characteristics of polymer coatings including drugs.

At least mild inflammation was found on the polymer plus drug stent and the polymer only stent. This finding indicates that the polymer shows good biocompatibility even without drug loading. Other studies have shown that when the drug is completely removed from the polymer, the polymer itself produces sufficient inflammation to cause neovascular intima. This observation may be responsible for the late catch-up phenomenon in the late stage of clinical restenosis. Because the polymer in this example did not cause inflammation in the coronary arteries, late-stage problems associated with polymers after drug depletion were unlikely.

In summary, the scaffold from the polymer-dissolved compound, quetiamus, showed a decrease in neovascular intimal hyperplasia when placed in the coronary artery in the porcine model.

Example 4

The purpose of this example was to determine the rate of release of the quetiamus drug from a biocompatible polymer coated 316L electropolished stainless steel coupon comprising a pendant phosphatidylcholine.

The rubber septum from the lid of the HPLC vial was removed from the vial and placed in a glass vial such that the "Teflon" side was facing up. These spacers are used as carriers for the test sample. The test sample was a 316L stainless steel test piece previously coated with a biocompatible polymer (PC polymer) comprising a side group of phosphocholine. Coronary stents are typically made of 316L stainless steel and can be coated with PC polymer to provide a reservoir for the loaded drug. The coated test piece for the simulated stent was placed on the septum. A solution of quetiamus and ethanol (10 μL) was applied to the surface of each test piece by using a glass Hamilton syringe. The solution included quetiamus (30.6 mg) dissolved in 100% ethanol (3.0 mL). The syringe was washed with ethanol between each application. Place the cap of the glass bottle loosely on the bottle to ensure proper ventilation. The test piece was allowed to dry for a minimum of 1.5 hours. Twelve coupons were loaded in this manner - six were used to determine the average amount of drug loaded onto the device, and six were used to measure the time required to release the drug from the device.

To determine the total amount of quetiamus loaded onto the test piece, the test piece was removed from the bottle and placed in a 50/50 acetonitrile/0.01 M phosphate buffer (pH 6.0, 5.0 mL). The test piece was placed in a 5210 Branson sonicator for one hour. The test piece was then removed from the solution and the solution was assayed by HPLC.

Fresh aliquots (10.0 mL) of 0.01 M phosphate buffer at pH 6.0 were immersed in and removed from each of the following time intervals - 5, 15, 30, and 60 minutes. Time release studies were performed. For the remaining time points of 120, 180, 240, 300, 360 minutes, a 5.0 mL volume of buffer was used. To facilitate mixing during drug release, the sample was placed on a low speed Eberbach shaker device. After the final sample test was completed, all solution aliquots were assayed by HPLC.

HPLC analysis was performed using a Hewlett Packard Series 1100 tool with the following configuration: injection volume = 100 μL acquisition time = 40 minutes flow rate = 1.0 mL / min column temperature = 40 ° C wavelength = 278 nm mobile phase = 65% acetonitrile / 35% H ₂ O column = YMC ODS-A S5 μm, 4.6 x 250 mm Part No. A12052546WT

The results of the above experiments show the release data of Table 6 below :

Example 5

The purpose of this example was to determine the loading and release of quetiamus from a 15 mm BiodivYsio drug delivery scaffold.

To load the stent with the drug, a solution of quetiamus in ethanol at a concentration of 50 mg/mL was prepared and dispersed in twelve bottles. The polymer coated twelve individual stents were placed on a fixture that was constructed to hold the stent in a vertical position and the stent was immersed vertically into the drug solution for five minutes. The stent and fixture are removed from the vial and the excess drug solution is aspirated by contacting the stent with the adsorbent material. The stent was then dried in air for 30 minutes in a reverse vertical position.

The scaffolds were removed from the fixture and each scaffold was placed in 50/50 acetonitrile/phosphate buffer (pH 5.1, 2.0 mL) and sonicated for one hour. The stent is removed from the solution and the drug concentration of the solution is assayed, which allows calculation of the initial amount of drug on the stent. This method is shown independently to remove at least 95% of the drug from the stent coating. The stent included an average of 120 ± 9 micrograms of drug.

The drug loaded scaffold was placed on the fixture in individual vials and placed in 0.01 M phosphate buffer (pH = 6.0, 1.9 mL). These samples were placed on a low speed Eberbach shaker device to provide back and forth agitation. To avoid approaching the saturation of the drug in the buffer, the scaffold was periodically transferred to fresh buffer bottles at 15, 30, 45, 60, 120, 135, 150, 165, 180, 240, 390 minutes. At the end of the drug release period studied, the drug concentration of the lysis buffer bottle was determined by HPLC. The data represents the cumulative release of the drug as a function of time, which is shown in tabular form in Table 7 below:

Example 6

The tetrazolium analog temazos of rapamycin has been shown to cause damage in porcine coronary stents (Touchard AG, Burke SE, Toner JL, Cromack K and Schwartz RS. Zotarolimus-eluting stents reduce experimental coronary artery neointimal hyperplasia After 4 weeks.Eur Heart J.27:988-993,2006).Delivered from the Biocompatibles BiodivYsio PC Coronary Stents in Porcine Coronary Arteries, Technical Report, Mayo Clinic and Foundation, Rochester, MN.) and Rat Balloon Angioplasty (Gregory, C. Summary of Study Evaluating Effects of zotarolimus in a Rat Model of Vascular Injury) The activity against restenosis in the model. The purpose of this example was to evaluate the safety and pharmacokinetics (PK) of a single intravenous (IV) dose of gradual increase of quetiamus in healthy men.

Currently, the first study in humans, the safety and pharmacokinetics of quetiamus, was studied after intravenous administration of quetiamus at a dose ranging from 100 to 900 μg. Intravenous single-dose administration mimics the fastest accidental release of sultasis from drug-coated stents in vivo.

This was a single escalating, double-blind, randomized, placebo-controlled, phase 1 single-center study. Sixty healthy adult males were divided into 5, IV, 100, 300, 500, 700, and 900 μg dose groups. The population information of the subjects is summarized in Table 8 .

Subjects were randomly assigned to receive a single intravenous dose of quetiamus or matched intravenous placebo in the fasted condition, as shown in the dosing diagrams shown in Table 9.

Higher doses were administered after assessing safety data from the aforementioned lower dose group. The treatment group was separated for at least 7 days. For safety reasons, each treatment component was divided into two groups of six subjects, and the doses of the two groups were separated by at least one day.

The dose was administered in a rapid IV over 3 minutes under 8 subjects. In each dose group, 4 subjects received quetiamus and 4 subjects received a placebo. Blood was sampled over 168 hours and the concentration of quetiamus was measured using LC-MS/MS with a LOQ of 0.20 ng/mL.

0.083 (5 min), 0.25, 0.5, 1, 2, 4, 8, 12, 16, 24, 36, 48, 72, 96, 120 before administration (0 hours) and after the first day of study At 144 and 168 hours, 7-mL blood samples were collected by venipuncture into a vacuum collection tube containing edetic acid (EDTA).

The blood concentration of quetiamus was determined using a confirmed liquid/liquid extraction HPLC tandem mass spectrometry method (LC-MS/MS). (Ji, QC, Reimer MT, El-Shourbagy, TA.: A 96-well liquid-liquid extraction HPLC-MS'/MS method for the quantitative determination of ABT-578 in human blood samples, J, of Chromatogr. B 805 , 67-75 (2004)). The lower limit of quantification of quetiamus using a 0.3 mL blood sample was 0.20 ng/mL. All calibration curves have a measurement coefficient (r ² ) greater than or equal to 0.9923.

Safety is assessed based on adverse events, physical examinations, vital signs, ECG, injection points, and laboratory test assessments.

Non-interval methods were used to assess the pharmacokinetic parameter values of quetiamus. These parameters include: quetiamus concentration (C ₅ ) at 5 minutes after administration, dose-normalized C ₅ , elimination rate constant (β), half-life (t _{1 / 2} ), from time 0 to final dose area of the blood concentration and the time curve in time concentrations measured (AUC _{0 - Finally),} dose-normalized of AUC _{0 - Finally,} extrapolated to the area under the curve of blood concentration versus time of infinite moment of (AUC _{0 - no limit)} , the dose normalized AUC _{0 - no limit,} total clearance (CL) and volume of distribution (Vd _β).

The mean blood concentration-time curves after intravenous administration of quetiamus are shown in Figures 4 and 5 , respectively, in a linear ratio and a logarithmic linear ratio.

The mean ± SD pharmacokinetic parameters of quetiamus are shown in Table 10 after administration of each of the two therapies.

$Harmonic mean ± false standard deviation; t _{1 / 2} evaluation based on statistical test of β; A> 10% of the subjects, samples of 201, 304 and 512 5 minutes sample sampling time deviation; such subjects The C ₅ concentration was not calculated. (N=7); # statistically important monotonic trend with dose.

To study the dose ratio and linear pharmacokinetics, covariance analysis (ANCOVA) was performed. Subjects were categorized by dose level and body weight was covariate. Analysis of the variables comprising _β, Vd β, the dose-normalized C ₅ and standardization of dose AUC _{0 -} standardization of _{the last} dose and AUC _{0 - no limit} on the number. The primary test that assumes a constant dose is a test of the dose level effect with respect to the monotonic function of the dose. In addition, the highest and lowest dose levels were compared within the framework of ANCOVA.

FIG Mok described Zhou Division _{C m a x, AUC 0 6} - _Finally and ₀ AUC _{- no limit} to the proportion of the dose into. Seen in this figure, a monotonic trend was not observed a statistically significant of which indicates that in these parameters of the dose-normalized C _{m a x} and AUC _{0 - Finally} increases proportionally to dose. About Ta Mok dose universe Secretary of standardized AUC _{0 - infinite,} monotonous trend observed significant dose of the concomitant significant (p = 0.0152). However, dose-normalized AUC ₀ for all groups _{- no limit} of the pairwise comparison shows only the 100 μg dose normalized AUC _{0 -} significantly different from the 900 μg and 300 μg of the dose-normalized AUC _{0 unlimited} statistically _{- infinite} (respectively p = 0.0032 and p = 0.0316.) A statistically significant monotonic trend with beta was also observed. This bias may be due to a slight overestimation of β in the 100 μg dose group. Zhou average Ta Mok Division C ₅ (a concentration at 5 minutes) and AUC _{0 - infinite} dose of a concomitant increases in proportion as shown in Table 11.

The mean half-life at the doses studied varied between 26.0 and 40.2 h and was not significantly different over the 300-900 μg dose range. Cytametol was well tolerated at all doses and no clinically significant physical examination results, vital signs or laboratory measurements were observed.

Safety The most common treatment-emergency adverse event associated with quetiamus (reported by two or more subjects in any treatment group) is the injection point response and pain.

Most adverse events are mild and naturally resolved to a severe extent.

There were no serious adverse events reported in this study.

There were no significant clinical changes in physical examination results, vital signs, clinical laboratory or ECG parameters during the study.

Conclusion On the 100-900 μg dose range of about C ₅ and AUC _{0 - unlimited} universe of IV Pharmacokinetic Mok Division dose proportional. In general, the 100-900 μg dose range in the universe Mok Division of substantially linear pharmacokinetics, such as by C _5, AUC _{0 - the last} and AUC _{0 - infinitely} increased proportional to the dose shown. A single dose of IV up to 900 μg was administered without consideration of safety.

The average elimination half-life of quetiamus varied from 26.0 to 40.2 hours over the range of doses studied. The average clearance and volume of the distribution varied from 2.90 to 3.55 L/h and 113 to 202 L, respectively. Regarding β and to a significant extent regarding Vd _β , the observed deviation from the linear kinetics was attributed to an overestimation of β for the 100 μg dose group.

Subjects are generally well tolerated with a single dose of quetiamus of 100 to 900 μg.

Example 7

This study was designed to assess the pharmacokinetics of quetiamus after multiple doses and to assess the safety of healthy subjects when their systemic exposure is maximized. The primary goal is to achieve a total exposure of quetiamus that is significantly higher than the expected level of drug eluted from the coated stent. The study investigated the pharmacokinetics and safety of quetiastat in a phase 1 and multiple dose escalation study (after multiple intravenous injections of 200, 400, and 800 μg per day for 14 consecutive days in healthy subjects). Sex.

Methods One-stage, multiple escalation doses, double-blind, placebo-controlled randomized studies. The 60-minute QD IV injection of quetiamus was divided into three daily once-a-day (QD) therapies (200, 400 or 800 μg QD, 16 of each active and 8 placebos) for 14 consecutive days. 72 subjects. Blood samples were collected 24 hours prior to dosing on the 10th, 11th, 12th, and 13th days after the first administration, and 168 hours after the 14th day. On days 1, 14, 16, 18 and 20, urine samples were collected over 24 hours. The concentration of quetiamus in blood and urine was determined using the confirmed LC/MS/MS method. The pharmacokinetic parameters were determined by interval analysis. All day AUC _{0 - ∞} (including all 14 doses from time 0 to infinite blood concentration-time curve area). Estimate dose and time - linear and stable state. The fraction of the drug eliminated in the urine is determined.

In this study, 72 generally healthy male and female subjects were enrolled. Population information is summarized in Table 12 .

As shown in Table 13 , subjects were randomly divided into three groups (Groups I, II, and III) at two different points. Within each group, subjects were equally divided between the two study sites, with 12 subjects enrolled at each site (8 subjects: quetamos, 4 subjects: placebo). The administration schedules in each dose group are shown below: + Subject 2112 stopped the study prematurely; on the 19th day of the study, the subject withdrew from the accreditation.

On Days 1 to 14 of the study, for Groups I, II, and III, subjects received a single 60-minute daily (QD) 200, 400, or 800 μg zeamolmus vein for fasting. Intravenous or placebo matched intravenous injection. The drug was administered via a syringe pump connected to a y-point device which was also injected with 125-150 mL of a 5% aqueous solution of dextrose (D5W) over 60 minutes. The group is then administered with the last dose of the previous group separated from the first dose of the next group for at least 7 days, during which time the safety data from the previous group is analyzed. The dose increase is dependent on the safety analysis of the lower dose group.

A 5-mL blood sample was collected in a tube containing potassium EDTA to evaluate 0.25, 0.5, 1.0, 1 hour, 5 minutes, 1.25, 1.5 after administration (0 hours) and on the first and 14th days of the study. , 2, 3, 4, 8, 12, 18 and 24 hours of quetiamus concentration. Additional samples were collected at the 36, 48, 72, 96, 120, 144 and 168 hours prior to the start of the study on day 14 of the study and on days 10, 11, 12 and 13. Urine was collected in preservative-free containers at the following intervals after the start of the study on days 1, 14, 16, 18 and 20: 0 to 6, 6 to 12, 12 to 18 and 18 to 24 hours.

The blood and urine concentrations of quetiamus were determined using a confirmed liquid/liquid extraction HPLC tandem mass spectrometry (LC-MS/MS). Using a 0.3 mL blood sample, the lower limit of quantitation of quetiamus was 0.20 ng/mL, and using a 0.3 mL urine sample, the lower limit of quantitation of quetiamus was 0.50 ng/mL.

Safety was assessed based on adverse events, physical examination, vital signs, ECG, injection location, and laboratory test evaluation.

Results The plasma concentration-time data of all subjects were described by a three-interval open model with first-order exclusion. In the treatments studied, the mean interval pharmacokinetic parameters ranged from: CL 4.0-4.6 L/h; V ₁ 11.3-13.1 L; V _{s s} 92.5-118.0 L and terminal exclusion t _{1 / 2} 24.7-31.0 h . On the first and the 14th day, the pharmacokinetics of quetiamus was consistent with the dose linearity on the treatments studied. The pharmacokinetic model met both day 1 and day 14 data, which is shown as time-linear pharmacokinetics. All the days of the treatment of the study AUC _{0 - ∞} 677-2395 ng. Hr/mL range. On average, an average of 0.1% of the amount of quetiamus can be recovered in the urine within 24 hours after administration.

Pharmacokinetics and Statistical Analysis The pharmacokinetic parameters of quetiamus were evaluated for individual subjects using interval analysis. Simultaneously simulating the first dose from the first day of the study, the last dose on the 14th day of the study, and the lowest concentration on the 10th, 11th, 12th, and 13th days of the study for each individual subject. data. The measured parameters are: volume of intermediate interval (V ₁ ), terminal exclusion rate constant (γ), clearance rate (CL), volume of distribution under steady state (V _{s s} ), half-life (t _{1 / 2} ), The maximum concentration (C _{m a x} ), the time of maximum concentration (T _{m a x} ), the area under the curve of blood concentration and time on day 14 (AUC _τ ), and the corresponding dose normalized C _{m a x} and AUC _τ . The best model of each individual subject was used to predict the concentration-time curve of individual subjects over a 14-day period to assess chronic exposure over the duration of the study, ie C _{m a x} and all-day AUC _{0 - ∞} ( Consider all 14 doses in the study, from 0 to infinity, the predicted area under the blood concentration-time curve).

To assess the dose proportionality of the study day 14 dose, covariance analysis (ANCOVA) was performed on dose normalized C _{m a x} , dose normalized AUC, and terminal elimination rate constants. The center and dose are factors and the body weight is a public variable. In order to solve the problem of whether or not the steady state is reached, the center and the dose level are used as factors to perform repeated measurement analysis on the dose pre-dose concentration normalized to the dose on days 10-14.

Pharmacokinetics The plasma concentration-time data of all subjects were described by a three-interval open model with first-order elimination. The mean blood concentrations of quetiamus on Day 1, Day 14, and Days 1 to 14 are shown in Figure 7. The mean ± SD of the pharmacokinetic parameters of quetiamus is shown in Table 14 .

$Harmonic mean ± false standard deviation ^* second prediction parameter

Because the deviation between the observed diagnostic curve and the predicted diagnostic curve was not observed on the therapy being studied, the range of interval pharmacokinetic parameters at the dose therapy studied was very narrow and not on the dose therapy studied. A meaningful trend on the second parameter was observed; the dose linearity of quetiamus was inferred on the dose therapy studied.

The following figure depicts the 14th day of quetiamus C_{m a x} And AUC_{0 - 2 4 h} It is proportional to the dose.Figures 8a, 8b and 8c Mean quetiamus blood concentration-time curves for the 200, 400, and 800 μg QD dose groups on Days 1, 14 and 1-14 were shown. For each dose group, the model fully describes the data on Days 1 and 14 and the days between them, such asFigure 9 Illustrated (examples of mean and predicted blood concentration versus time curves for 800 μg QD dose group data). The excellent quetiamus concentration-time data observed by day 3 to day 14 by assuming a linear dynamics 3 interval model is consistent with the quetiamus display time constant clearance rate.

As shown in FIG. 9, 10-14 days on study dose-normalized pre-dose concentrations not observed statistically significant.

The median values of C _{m a x} for the 200, 400, and 800 μg QD dose groups were 11.4, 22.1, and 38.9 ng/mL, respectively. Corresponding to the full-day AUC _{0 - ∞} median values are 677, 1438 and 2395 ng. h/mL.

The fraction of the temasis dose eliminated in the urine was calculated for the 800 μg QD dose group. On average, about 0.1% of the quetiamus was recovered in the urine within 24 hours on the first day and the 14th day.

Safety The most common treatments associated with quetiamus - emergency adverse events are pain, headache, injection point reaction, dry skin disease, abdominal pain, diarrhea and rash. Most adverse events are mild and naturally resolved to a severe extent. There were no serious adverse events reported in this study. In particular, none of the subjects showed any clinical or biochemical evidence of immunosuppression, delayed QTc, or clinically significant adverse events.

Conclusions In the dose therapy studied, the pharmacokinetics of quetiamus was proportional to the dose and constant over time when administered intravenously for 14 consecutive days.

The steady state of QD administration of quetiamus was reached on day 10, on which the first lowest sample was measured.

Because about 0.1% of the daily dose is excreted in the urine as an unaltered drug, renal excretion is not the primary route of elimination of quetiamus.

Cystatol is generally well tolerated when administered 200, 400 and 800 μg multiple times for 14 consecutive days.

Example 8 Individual anti-inflammatory effects of quetiamus and dexamethasone

Percutaneous transluminal coronary angioplasty (PTCA) and post-stent inflammatory response have been shown to play an important role in vascular remodeling associated with restenosis{RGMacdonald, RSPanush and CJPepine, Rationale for use of glucocorticoids in modification Of restenosis after percutaneous transluminal coronary angioplasty, Am J Cardiol, 60, 3, 1987, 56B-60B; JS Forrester, M. Fishbein, R. Helfant and J. Fagin, A paradigm for restenosis based on cell biology: clues for the development Of new preventive therapies, J Am Coll Cardiol, 17, 3, 1991, 758-69; SPKaras, EC Santolian and MB Gravanis, RestenoSis following coronary angioplasty, Clin Cardiol, 14, 10, 1991, 791-801; P. Iibby And SK Clinton, Cytokines as mediators of vascular pathology, Nouv Rev Fr Hematol, 34 Supplement, 1992, S47-53}. A large number of cell types promote this response, including circulating monocytes, macrophages, neutrophils, eosinophils, platelets, vascular smooth muscle and endothelial cells {RGMacdonald, RSPanush and CJPepine, Rationale for use of glucocorticoids in Modification of restenosis after percutaneous transluminal coronary angioplasty, Am J Cardiol, 60, 3, 1987, 56B-60B}. Activation of immune cells following PTCA results in cytokine/chemokine production and recruitment of circulating mononuclear cells to the vessel wall. The three important cytokines produced by monocytes are tumor necrosis factor (TNFα), monocyte chemoattractant protein 1 (MCP-1), and interleukin 6 (IL-6). It has been shown that it increases with PTCA and the increased levels of MCP-1 and IL-6 are associated with an increased incidence of restenosis {Y.Hojo, U.Ikeda, T.Katsuki, O.Mizuno, H.Fukazawa, H.Fujikawa and K. Shimada, Chemokine expression in coronary circulation after coronary angioplasty as a prognostic factor for restenosis, Atherosclerosis, 156, 1, 2001, 165-70; F. Cipollone, M. Marini, M. Fazia, B. Pini, A. Iezzi , M.Reale, L.Paloscia, G.Materazzo, E.D'Annunzio, P.Conti, F.Chiarelli, F.Cuccurullo and A.Mezzetti,Elevated circulating levels of monocyte chemoattractant protein-1 in patients with restenosis after coronary Angioplasty, Arterioscler Thromb Vase Biol, 21, 3, 2001, 327-34}. Tumor necrosis factor (TNFα) is an inflammatory cytokine produced by monocytes and tissue macrophages (cell types already suggested in the pathophysiology of restenosis) (Moreno PR, Bernardi VH, Lopez-Cuellar J, Newell JB, McMellon) C, Gold HK, Palacios IF, Fuster V, Fallon JT. Macrophage in filtration predicts restenosis after coronary intervention in patients with unstable angina. Circ, 94 (12), 1996: 3098-102). It has been shown that TNFα levels are increased in humans with in-stent restenosis and this cytokine content is increased after angioplasty (Kozinski M, Krzewina-Kowalska A, Kubica J, Zbikowska-Gotz M, Dymek G, Piasecki R, Sukiennik A, Grzesk G, Bogdan M, Chojnicki M, Dziedziczko A, Sypniewska G. Percutaneous coronary intervention triggers a systemic inflammatory response in patients treated for in-stent restenosis-comparison with stable and unstable angina:Inflammation Research:54:2005:187- 93). It has been shown that anti-TNFα antibodies loaded onto the scaffold reduce PCNA (a marker of cell proliferation in human saphenous vein culture), suggesting that blocking TNFα action results in a decrease in neovascular intimal hyperplasia (Javed Q, Swanson N, Vohra H, Thurston H, Gershlick AH.; Tumor necrosis factor-alpha antibody eluting stents reduce vascular smooth muscle cell proliferation in saphenous vein organ culture. Exp Mol Path, 73(2): 2002: 104-11). In addition, increased levels of TNFα stimulate vascular smooth muscle and endothelial cells to produce cytokines and this can lead to sustained inflammatory responses in the vessel wall.

The glucoside dexamethasone has an effective anti-inflammatory effect on a large number of cell types. Dexamethasone has been proposed for use as an anti-restenosis agent and is currently used on Dexamet drug dissolving scaffolds (available in Europe from Abbott Vascular Inc). (Check) {D.W. Muller, G. Golomb, D. Gordon and R. J. Levy, Site-specific dexamethasone delivery for the prevention of neointimal thickening after vascular stent implantation, Coron Artery Dis, 5, 5, 1994, 435-42}. However, the previously unknown immunosuppressive agents, temasimus and sirolimus, have an effect on the production of these important cytokines by human monocytes.

In other embodiments, the second drug is a member of a group consisting of small molecules and biological agents that reduce the activity of inflammatory cytokines. In other embodiments, the second medicament comprises an anti-TNFα therapeutic agent consisting of adalimumab, an anti-MCP-1 therapeutic agent and a CCR2 receptor antagonist, an anti-IL-18 therapeutic agent, an anti-IL-1 therapeutic agent and Salts, esters, prodrugs and derivatives or any combination thereof.

Experiments were performed to determine the effects of quetiacin, sirolimus, and dexamethasone on the production of TNFα, IL-6, and MCP-1 by activated human monocytes. In addition to monocytes and macrophages, two other important cell types are involved in the etiology of restenosis, arterial smooth muscle, and endothelial cells. These cells are activated in response to TNFα and produce cytokines. To determine the anti-inflammatory effects of dexamethasone, quetiacin and sirolimus, human coronary endothelial and smooth muscle cells (hCaEC and hCaSMC, respectively) were tested. The activation of cytokines produced by human coronary artery smooth muscle and endothelial cells after TNFα treatment was measured by measuring the production of interleukin-8 (IL-8), MCP-1 and IL-6. As shown by the experimental results described below, the ability of these compounds to inhibit the production of cytokines supports their use as anti-restenosis agents in combination because these cytokines contribute to the development of restenosis.

Methods Human monocytes (Cambrex, East Rutherford NJ) were inoculated into a 96-well microtiter plate (60,000 cells per well), incubated at 37 ° C for 48 hours, and then used in the presence or absence of test compounds in the culture medium. Lipopolysaccharide (LPS, 25 or 100 ng/ml) was stimulated for 24 hours. After 24 hours, the supernatant was carefully collected and the TNFα, IL-6 and MCP-1 contents were determined by ELISA. Major cultures of human coronary endothelium and smooth muscle cells were obtained from Cambrex and maintained as described by the merchant. The cells were seeded into 96-well microtiter plates at a concentration of 5000 cells per well (hCaSMC) and 7500 cells per well (hCaEC) and allowed to attach. After 24 hours, the supernatant was removed and replaced with medium containing TNFα (5 ng/mL) in the presence or absence of various concentrations of quetiamus, dexamethasone or sirolimus. The cells were incubated for 24 hours and the supernatant was carefully removed and frozen until measurement. Cytokine levels were measured by sandwich ELISA in the supernatant.

Results and Conclusions Figure 10 shows that dexamethasone, quetiacin and rapamycin both dose-dependently inhibit the production of MCP-1 by human monocytes in vitro. However, quetiamus and rapamycin are more potent and effective than dexamethasone in this effect. Conversely, dexamethasone effectively blocked the production of TNFα and IL-6, and quetiamus or rapamycin did not demonstrate these effects ( Figures 10 and 11 ). In hCaSMC, the agent demonstrating activity was only dexamethasone, which inhibited cytokine production on a dose-dependent basis. Dexamethasone blocked 57.2% of hCaSMC MCP-1, 65.7% of IL-6 and 68.4% of IL-8. Information on all three cell types is summarized in Table 15 . None of the tested agents inhibited cytokine production of hCaEC by 50% compared to the control group. Because the blockade of all three cytokines (TNFα, MCP-1, and IL-6) has a beneficial anti-inflammatory effect on preventing restenosis, this data supports the combined use of quetiamus and dexamethasone.

Example 9 Antiproliferative activity of quetiamus and dexamethasone

Dexamethasone has been proposed for use as an anti-restenosis agent and is currently used on dexamethasone drug-dissolving stents (available in Europe from Abbott Vascular Devices). Dexamethasone has been previously shown to be an effective hCaSMC rather than a hCaEC anti-proliferative agent (Li L, Burke, SE, Chen, Y-CJ. Comparison of drugs in inhibiting human smooth muscle and endothelial cell proliferation.: Abbott Laboratories Corporate Technology Exchange Poster Presentations; October 27, 2003: R. Voisard, U. Seitzer, R. Baur, PC Dartsch, H. Osterhues, M. Hoher and V. Hombach, Corticosteroid agents inhibit proliferation of smooth muscle cells from human atherosclerotic arteries in vitro, Int J Cardiol, 43, 3, 1994, 257-67). In addition to its anti-proliferative activity, dexamethasone has an effective anti-inflammatory effect. Inflammation after stent implantation has been suggested to promote restenosis, and agents that interfere with vascular inflammatory responses can attenuate restenosis (P. Libby and SK Clinton, Cytokines as mediators of vascular pathology, Nouv Rev Fr Hematol, 34 Supplement, 1992, S47 -53).

Experiments were performed to study the interaction between quetiamus and the corticosteroid dexamethasone. The in vitro proliferation assay was used to determine the effect of dexamethasone on the antiproliferative activity of quetiamus in human coronary artery smooth muscle (hCaSMC) and endothelial cells (hCaEC). Proliferation of vascular smooth muscle cells and migration to the vascular neovascular intima is a characteristic pathological response seen in restenosis lesions (A. Lafont and P. Libby, The smooth muscle cell: sinner or saint in restenosis and the acute coronary syndromes? J Am Coll Cardiol, 32, 1, 1998, 283-5). As a result, in vitro assays that specifically measure the anti-proliferative activity of candidate anti-restenotic compounds against human coronary artery smooth muscle and endothelial cells should predict potential anti-restenotic activity in vivo. The 氚 incorporation assay is an accurate and sensitive method for determining cell number and proliferation in this technique. Compounds or compounds that attenuate growth factor-regulated human coronary artery smooth muscle cell (hCaSMC) proliferation are candidates for candidate anti-restenosis agents as measured by the in vitro sputum incorporation assay. This assay was used to determine whether an agent exhibiting anti-proliferative activity in a single case also showed similar activity in the combination.

In addition, agents that exhibit inefficient antiproliferative activity block the activity of more potent anti-proliferative agents when administered in combination. A clear example of the effect of tacrolimus on the weakening of the antiproliferative activity of quetiamus ( Fig. 12 ). As can be seen, tacrolimus ("T") blocks the antiproliferative activity of quetiamus in hCaSMC. Tacrolimus (100 nM) to increase the universe of IC _{5 0} Division Mok and tacrolimus (250 nM or greater) completely blocked the universe Mok Division (4 nM) of the activity. Although the combination of each affects different signal pathways, both quetiamus and tacrolimus bind to the co-receptor FKBP-12.

To determine the potential anti-restenotic activity of the combination of quetiamus and dexamethasone, the proliferation of hCaSMC and hCaEC was measured in the presence of each individual compound and combination.

Dexamethasone inhibits the synthesis of important proteins required for cell division in some cell types and is an effective anti-inflammatory drug (Voisard R, Seitzer U, BaurR et al. Corticosteroid agents inhibit proliferation of smooth muscle cells from human atherosclerotic arteries in vitro. Int J Cardiol. March 1, 1994; 43(3): 257-267; N. Baghdassarian, A. Peiretti, E. Devaux, PA Bryon and M. French, Involvement of p27Kip1 in the G1-and S/G2-phase lengthening mediated by Glucocoliticoids in normal human lymphocytes, Cell Growth Differ, 10, 6, 1999, 405-12). Similar to rapamycin, quetiamus blocks cyclin-dependent kinase via mTOR inhibition and inhibits cell cycle progression in the G1-S phase (SOMarx, T.Jayaraman, LOGO and ARMarks, Rapamycin-FKBP inhibits cell cycle regulators) Of proliferation in vascular smooth muscle cells, Ore Res, 76, 3, 1995, 412-7; Sehgal SN, Rapamune (RAPA, rapamycin, sirolimus): mechanism of action immunosuppressive effect results from blockade of signal transduction and inhibition of cell cycle progression Clin Biochem. July 1998; 31(5): 335-340; SNSehgal, Sirolimus: its discovery, biological properties, and mechanism of action, Transplant Proc, 35, 3 Supplement, 2003, 7S-14S). To determine whether dexamethasone attenuates or increases the antiproliferative activity of quetiamus on hCaSMC, the effect of these agents, alone or in combination, on proliferation induced by growth factors is determined. Since re-endothelialization of vascular injury is considered to be beneficial, the anti-proliferative effects of the two agents on hCaEC were evaluated, alone and in combination. Information on interactions (increases) was analyzed using equivalent dose methods and combination index analysis as described below.

Proliferation assay ³ H thymidine uptake assay Cell proliferation was monitored by incorporation of ³ H-thymidine into newly synthesized cellular DNA (stimulated by serum and growth factors). The exponentially growing hCaSMCs were seeded in 96-well flat-bottom tissue culture dishes at 5000 cells per well (10,000 cells per well for hCaEC). The cells were attached overnight. The next day, the growth medium was removed and the cells were washed twice with unsupplemented (basal) medium to remove traces of serum and growth factors. The basal medium (200 μL) was added to each well and the cells were incubated in a medium lacking growth factors and serum to starve and synchronize them to the G ₀ state. After starvation in the medium lacking serum and growth factors (48 hours for hCaSMC and 39 hours for hCaEC), the cells were supplemented with 200 μL of supplemented medium in the absence or presence of the desired concentration of the drug. DMSO was maintained at a final concentration of 0.1% in all wells. After a 72-hour incubation period, 25 μL (1 μCi per well) of ³ H-thymidine (Amersham Biosciences) was added to each cell. The cells were incubated at 37 °C for 16-18 hours to incorporate ³ H-thymidine into the newly synthesized DNA and harvested into 96 wells containing the bonded glass fiber filter using a cell harvester (Harvester 9600, TOMTEC). On the plate. Filter plates were air dried overnight and MicroScint-20 (25 μL) was added to each filter well and the plates were counted using a TopCount micro-quantitative disk scintillation counter. The control group included only medium, starved cells, and cells in intact medium.

The drug activity was established by measuring the inhibition of the incorporation of ³ H-thymidine into the newly synthesized DNA relative to cells grown in intact medium.

Data are presented as percent inhibition of ³ H-thymidine incorporation relative to vehicle-treated controls and are mean ± SEM of 3-4 experiments. Generating a semi-logarithmic curve from the average value of inhibition of the experimental drug concentration, and by cultivating in respect of the complete medium lacking extracellular medicament push content 50% inhibition of each experiment to determine the IC _50. Final IC _{5 0} of the experiment the average of 3-4.

In these experiments, the X axis represents the varying drug concentration. For most of these curves, it is a single quetiamus and a combination of quetiamus with a fixed concentration of dexamethasone. Each plot contains a separate dexamethasone curve and a separate quetiamus curve. The curves in each figure were set by adding dexamethasone (1, 5, 10 or 25 nM) to various concentrations of quetiamus (0, 0.04, 0.08, 0.8, 4, 8, 40 nM) at a fixed concentration. To produce. Each curve represents a dose response of quetiamus (the concentration of which is given on the X-axis) in the presence of a fixed concentration of dexamethasone (given in the legend). It should be noted that higher fixed concentrations of dexamethasone (1, 10, 100, 1000 nM) are used in hCaEC.

Two methods were used to analyze the effect of quetiamus and dexamethasone combination on hCaSMC proliferation. The Tallarida method was used at several effective levels to generate an isobologram (RJ Tallarida, Drug synergism: its detection and applications, J Pharmacol Exp Ther, 298, 3, 2001, 865-72). A concentration response curve conforming to the nonlinear regression (Prism, GraphPad) to obtain EC _{5 0} and the gradient value. The 4-parameter equation was used to determine the concentration that caused the specific anti-proliferative effect (Equation 1) Y = bottom + (top-bottom) / (1 + 10 ^ ((LogEC _{5 0} -X) * slope))

X is the logarithm of the concentration. Y is the reaction or:

Where X = the logarithm of the drug concentration that produces the Y reaction, and the top and bottom values are limited to 100 and 0, respectively. In addition to the isobologram, the Chou and Talala methods were used to coordinate the analysis with the following exceptions (TCChou and P. Talalay, Quantitative analysis of dose-effect relationships: the combined effects of multiple drugs or enzyme inhibitors, Adv Enzyme Regul, 22 , 1984, 27-55). The regression model generated for each curve was replaced by a median impact data (log-logit curve), which is a more accurate simulation of the concentration-response curve because of the nonlinear 4-parameter equation. The median impact curve is severely affected by fractional occupancy values below 0.2 and above 0.8. Calculating the combination index (CI) of 25%, 50%, 60%, and 75% of several drug combinations according to Equation 2 (Chou TC, Talalay P. Quantitative analysis of dose-effect relationships: the combined effects of multiple drugs or enzyme Inhibitors. Adv Enzyme Regul. 1984;22:27-55).

(D) ₁ /(D _x ) ₁ +(D) ₂ /(D _x ) ₂ +((D) ₁ (D) ₂ )/(D _x ) ₁ (D _x ) ₂ =CI (Equation 2)

Wherein (D) ₁ and (D) ₂ are the concentrations of the drug 1 and the drug 2 in the combination, and (D _X ) ₁ and (D _x ) ₂ are the concentrations of the drug 1 alone and the drug 2 alone.

Assuming that each drug acts according to its potency, the CI value reflects the sum of the effects of the combination. Equation 2 describes the predictive effect of a combination of two non-specific compounds with each other. CI is equal to 1 if each drug promotes a combined effect based on its own dose-dependent fractional occupancy. Since CI depends on the extent of the observed effect, CI is determined at several levels of action using multiple drug combinations. The CI value is plotted as a function of the degree of action (or f _a ) (the CI value is calculated below). Similar to the isobologram analysis method, the CI value depends on the degree of action and varies with the degree of action. Therefore, it is important to consider the degree of action when comparing CI values. The accuracy of the CI value is then determined by the accuracy of the concentration values used in its calculations. In this study, a precise method (in accordance with the asymptotic curve of the GraphPad software) was used to calculate the drug concentration from each cumulative dose response curve for several degrees of action. The dose response curve can be in accordance with the data, which demonstrates activity that is almost dose independent. This is especially evident when analyzing the dose response curve produced in the presence of one of the high concentration test agents. In such cases, an error in the concentration of the drug determined from the dose response curve can result in _a high CI value at _a low degree of action (f _a ). Thus, a well-defined dose response curve generated near or above the half-value-maximum effect (i.e., f _a about 0.5) produces a CI value that is the most accurate predictor of drug combination activity. In these cases, it is considered that the CI value below 1 is superadded, and the value significantly exceeding 1 is weakly increased. It is considered that the value close to 1 is forgiveness.

Results and Conclusion Figure 13 shows dexamethasone, but not affect the proliferation of hCaEC blocked hCaSMC dose-dependent proliferation, wherein the IC _{5 0} to 9.3 nM (Table 16). In addition, the data show that the presence of dexamethasone does not impair the anti-proliferative activity of quetiamus, that is, the combination of these drugs is complementary. The data ( Figure 13 ) also showed that increasing the concentration of dexamethasone and quetiamus resulted in significant inhibition of hCaSMC cell proliferation. Because the combination shows activity in vitro, these data predict that the combination of quetiamus and dexamethasone is beneficial in preventing restenosis.

Higher dexamethasone doses achieve a higher degree of proliferative inhibition. The equivalent dose analysis of hCaSMC is given in Figure 14 , and the combination index (CI) analysis is Figure 15 . In Figure 14 , the concentration at which a particular degree of activity is produced is determined from a dose response curve generated by a non-linear curve consistent with the mean of the data. In Figure 15 , CI levels were determined from the mean data using the Chou and Talalay methods referenced above. f _a represents fractional percent inhibition.

Equivalent line graph analysis was performed at various levels of action to determine whether the combination of quetiamus and dexamethasone has potentiating activity in inhibiting cell proliferation. Although the combined index analysis shows similar results, all degrees of action can be shown in a graph. Both the isobologram and the combination index analysis showed that the combination of quetiamus and dexamethasone blocked the proliferation, and because the CI value was close to 1 at the half-maximum effect level, the effect was similar to the total individual The role predicted by the action of the agent. The combined analysis of the data from hCaEC was not performed because dexamethasone did not demonstrate activity in this cell type.

Although dexamethasone blocked hCaSMC proliferation, it did not affect hCaEC proliferation. In addition, because it is believed that re-endothelialization of the injured vessel wall is beneficial for preventing restenosis, such data support the use of dexamethasone as an anti-restenosis agent.

Example 10 dissolution test

I. Coating the stent with PC-1036 (Whether PC-2126 should be added? John) The coated stent was prepared prior to any experiment. These brackets are 3.0 mm x 15 mm 316 L electropolished stainless steel brackets. Each clean scaffold was sprayed with a solution of 20 mg/mL phosphonium choline polymer PC-1036 (product of Biocompatibles Ltd., Farnham, Surrey, UK) in ethanol. The stent was initially air dried and then cured at 70 ° C for 16 hours. It is then subjected to gamma irradiation at less than 25 KGy.

II. Using the drug-loaded stent of interest In these experiments, the agent was loaded onto the stent and the dissolution profile was examined. In general, the process is as follows. A plurality of PC coated stents are loaded with a drug or drug combination solution. The drug solution is typically in the range of 2-20 mg/mL of quetiamus and 10.0 mg/mL of dexamethasone in 100% ethanol, wherein about 10% of PC-1036 is added to the solution to enhance film formation. Loading the drug combination and individual drugs onto the stent is accomplished by spraying the desired drug spray onto the stent in a one-way spray system in a separator unit. All drug-dissolving stents were fabricated from Abbott's proprietary TriMaxx construction 15 mm x 3.0 mm stents, and all catheters were Medtronic (Minneapolis, MN) OTW, 15 mm x 3.0 mm. The quantities manufactured for each combination provide sufficient quantities for accelerated dissolution, drug loading levels, impurity profiles, and animal efficiency tests.

The scaffold is weighed and then loaded with the drug solution. All scaffolds were spray-loaded to their target drug content from a solution containing the appropriate drug and PC1036 in ethanol (in a ratio of 91:9). For the dexamethasone: quetiamus combination, the stent was prepared with each drug at 10 μg/mm. 10 μg/mm of individual quetiamus was loaded onto a single drug stent. Once loaded, all of the stents were dried in an open bottle at 40 ° C for 30 minutes in an open bottle and weighed to determine the drug load. The drug-loaded scaffold was then overcoated with 5 μg/mL PC1036 by spraying with a 10 mg/mL polymer in ethanol.

After the overcoating, the stent was cured in an oven at 70 ° C for 2 hours, and then weighed to determine the weight of the overcoat layer. After the drug is loaded, the stent is assembled onto the catheter and crimped onto the balloon. The coating and physical defects of the stent are then visually inspected. Each stent/catheter is inserted into a package ring and then a Tyvek bag. The bag was sealed with a Vertrod instant heat sealer. Place the bracket identification mark in the bottom corner on the front side of the bag, outside the sealed area containing the product. The product is then placed in a white box that is in detail labeled with the product and shipped for EtO sterilization. After sterilization, the product is packaged in a foil pouch containing an oxygen scavenger and a desiccant capsule. Mark the bag with the bracket identification number and product details. The bag was packaged while flushing with nitrogen.

III. Extraction of Drugs from Scaffolds For each drug or combination of drugs tested, three scaffolds were used to assess the total amount of drug loaded by the above method. The stent was immersed in 6 mL of 50% acetonitrile, 50% buffer solution and sonicated for 20 minutes. The concentration of each drug in the extraction solution was analyzed by HPLC.

At the end of the dissolution experiment discussed below, the scaffold was removed from the dissolving medium and immersed in 6 mL of 50% acetonitrile, 50% buffer solution and sonicated for 20 minutes. The concentration of each drug in these bottles indicates the amount of drug remaining on the stent at the end of the dissolution test.

IV. Dissolution Procedure For the evaluation of in vitro drug detachment, the stent (n=3 of each group) was developed and then placed in a solution of 10% mM acetate buffer (pH=4.0) with 1% Solutol HS 15 (in USP) Heated to 37 ° C in a Type II dissolution apparatus. Solubilizers are needed because of the extremely low water solubility of the drugs. The buffered medium is used to minimize degradation of the "Moss" drug (which occurs at pH values above 6). Treatment with buffer at pH 4 solves this problem. Because these drugs have minimal dissociation in these pH ranges, pH has little effect on the dissolution rate.

Samples were taken from the dissociation tank using an injection sampler equipped with only Teflon, stainless steel or glass surfaces at selected time intervals. Aliquots were collected after 15 min, 30 min, 1 h, 2 h, 4 h, 6 h, 8 h, 12 h, 18 h and 24 h. Samples were analyzed for concentration of quetiamus and dexamethasone via HPLC. Data are expressed as the average percentage of dissolved drug and dissolved in micrograms.

In the HPLC method, column switching is required to minimize the Solutol contamination of the analytical column and allow for pre-washing of the catheter column, or the system is coated with Solutol and the chromatographic retention is significantly altered. The sample is first injected into the front catheter column. Once the analyte peak is eluted from the anterior catheter column and passed through the analytical column, the anterior catheter column is converted out of the analytical path. The pre-catheter column is then washed to remove Solutol prior to the next injection.

V. Dissolution results Figures 15, 16 and 17 show loadings with 10 μg/mm quetiasti and dexamethasone (loads of each drug to a polymer PC-1036 overcoat with 5 μg/mm as described above) The accelerated dissolution rate of the stent.

In Figure 15 (dissolved drug in micrograms), the 24 hour dissolution profile shown is for the combination of the anti-inflammatory drug dexamethasone and the anti-proliferative agent quetiamus. The dissolution was carried out as described above. Although dexamethasone initially had a slightly greater burst than quetiamus, the two drugs were subsequently released at almost the same rate.

Figure 16 (% of dissolved drug) depicts the data in Figure 15 of the total drug standard determined on the stent after final stent extraction. As can be seen, the two drugs were removed 100% from the stent coating. The total drug recovered is very consistent with the drug load predicted by the weight gain of the stent during the drug loading process. This information and the drug efficacy of the same batch and the relevant substance test on the stent indicate that the drug is stable in the polymer coating when manufactured as described above. Small error bars (representing standard deviation) indicate that a dual drug lysate scaffold can be fabricated under reproducible dissolution kinetics.

In Figure 17 , the two curves are the dissolution curves of the individual quetiamus and the quetiamus in the presence of dexamethasone (micrograms of release versus time) under the same conditions. As can be seen, the two curves are very similar, indicating that dexamethasone has little effect on the dissolution curve of quetiamus.

Example 11 Test of neovascular intimal hyperplasia and endothelialization after stent implantation

This test was used to determine the effect of dual drugs on neovascular intimal hyperplasia and endothelialization. The test uses a technically recognized porcine coronary over-extension model (Schwartz, RS, Restenosis and the proportional neointimal response to coronary artery injury: results in a porcine model. J Am Coll Cardiol. February 1992; 19(2): 267 -274), and usually for about 2-8 weeks. Typically the experimental construct comprises at least one scaffold control that is completely similar to the experimental scaffold except that it changes a single variable, including the therapeutic substance or polymer.

In one example, a test scaffold was used in each pig (Cypher And Taxus Two major coronary arteries were implanted each, and a third primary coronary artery was implanted with a control ZoMaxx ^{T M} stent. Additional examples include implanting three TriMaxx A control stent was implanted in each of the major coronary arteries. The response of these control scaffolds can be compared with ZoMaxx ^{T M} and Cypher from self-implanted animals. And Taxus The responses obtained by the stent were compared.

The stent is implanted using standard techniques. At the end of the study, animals were euthanized and the hearts removed, washed and fixed using standard tissue antiseptic techniques (including formalin, formaldehyde, etc.). The stretched blood vessels are excised and then infiltrated and implanted into a suitable medium (including methyl methacrylate (MMA), paraffin or cryogenic medium) for sectioning. Divide all segments containing the stretched blood vessels to obtain informational information: (eg) three stents and two control sections. Continuous thin sections (approximately 5 μm) are typically used to various extents and stained to visualize cells and tissues (eg, hematoxylin and eosin (HE) and Masson's Verhoeff Elastin (MVE)). Slices are assessed and evaluated using image analysis systems or other technically accepted methods of morphological data collection and quantification. Data were evaluated for neovascular intimal area, neovascular intima thickness, and percentage-stenosis area.

Schwartz, RS, Restenosis and the proportional neointimal response to coronary artery injury: results in a porcine model. J Am Coll Cardiol. February 1992; 19(2): 267-274) The formation of neovascular intima was examined 28 days after stent implantation. The study evaluated a large number of randomized drug-eluting stents and a drug-coated drug-coated ZoMaxx ^{T M} stent. In each pig, two primary coronary arteries were implanted with one test stent and the third major coronary artery was implanted with a ZoMaxx ^{T M} stent. The ZoMaxx ^{T M} stent includes 10 μg/mm quetiamus as an active pharmaceutical agent. In addition, with three bare metal TriMaxx The stents were each implanted in three pigs (9 stents in total) for comparison. For the purposes of this disclosure, compare the five types of stents: 1) ZoMaxx ^{T M} stent (3.0 x 15 mm) containing 10 mcg of quetiamus per millimeter; 2) commercially available 8.5 mcg sirolimus per millimeter Cytox-polymer coated Cypher (3.0 x 13 mm) scaffold (as defined in the Cordis FDA statement); 3) commercially available Pacific paclitaxel-polymer coated Taxus per mile including 6.8 mcg of paclitaxel (calculated) (3.0 x 16 mm) scaffold; 4) Stent coated with a combination of quetiamus and dexamethasone (3.0 x 15 mm, coated with 10 mcg of each drug per mm); 5) TriMaxx excluding drug Bracket (3.0 x 15 mm).

The stent was implanted using a 1.30 balloon/artery ratio as determined by quantitative coronary angiography.

There were no deaths associated with the heart or stent during the study. After 28 days, the animals were euthanized and the hearts were removed and fixedly perfused with a lactated Ringer solution at approximately 100 mm Hg until blood clearance followed by fixation with 10% neutral buffered formalin. The expanded vessel is then excised, then infiltrated and implanted with methyl methacrylate (MMA). Sections containing all stretched blood vessels were sectioned with sections containing three scaffolds plus two control sections. Two consecutive thin sections (about 5 microns) were used to varying degrees and stained with hematoxylin and eosin (HE) and Masson's Verhoeff Elastin (MVE). The sections were evaluated and evaluated using the BIOQUANT ^{T M} TCW98 image analysis system. The mean values of neovascular intimal area, neovascular intima thickness, and stenotic area % are shown in Figures 18-20 for all stents in the five groups.

With TriMaxx Compared to the stent, as measured by conventional morphometric measurements, ZoMaxx ^{T M} , Cypher And Taxus The stent similarly reduces the formation of neovascular intima. With TriMaxx Compared to stents, stents including the combination of quetiamus/dexamethasone also showed a significant reduction in neovascular intimal hyperplasia. In addition, with zetamoli-polymer coated ZoMaxx ^{T M} stent, sirolimus-polymer coated Cypher Stent and Pacific Paclitaxel-Polymer coated Taxus Such quetiamus/dexamethasone combination stents ("Zot/Dex 10/10") also showed a further improvement in neovascular intimal reduction compared to stents.

Table 17 summarizes the polymer coated ZoMaxx ^{T M} stent and the quetiamus/dexamethasone combination drug stent with TriMaxx as a reference The improvement achieved by the stent compared to the stent.

With TriMaxx Compared with the control stent, the prior art single drug stent ZoMaxx ^{T M} , Cypher And Taxus Each showed a significant reduction in neovascular intimal formation. For example, the neovascular intima of the ZoMaxx ^{T M} stent was reduced by an average of 34.5% compared to the control group. The stent with 10 mcg of quetiamus/10 mcg dexamethasone combination ("Zot/Dex 10/10") per millimeter is even further reduced than the impressive results seen under the best single drug stents in commercial and clinical trials. Neovascular intima formation. When working with TriMaxx The quetiamus/dexamethasone combination drug-dissociated stent reduced the neovascular intima formation by an average of 49.3% when compared to the drug-free stent. With ZoMaxx ^{T M} and Cypher And Taxus The additional significant reductions in neovascular intimal hyperplasia were 22.6%, 25.4%, and 25.2%, respectively, compared to drug-eluting stents (Table 18).

Based on previous information published in the literature (Suziki, T et al, Stent-based delivery of sirolimus reduces neointimal formation in a porcine coronary model. Circulation. 2001; 104: 1188-1193) and patent application (Falotico R US patent application) 2003/0216699; Table 6.0), the skilled artisan concluded that the combination of Moss and dexamethasone lacks any anti-proliferative benefit over either drug alone. In fact, Table 6.0 clearly describes this effect. In contrast, we have unexpectedly demonstrated that the quetiamus/dexamethasone combination scaffold is highly efficient, which improves the reduction of neovascular intimal hyperplasia in the widely used porcine coronary artery overextension model. In one embodiment, the therapeutic amount of the Moss drug comprises quetiamus or everolimus and is at least 1 μg per millimeter of stent. In another embodiment, the second drug is dexamethasone and the therapeutic amount is at least 0.5 μg per millimeter of stent.

Figures 21 to 24 demonstrate the significant difference between the results of quetiamus and dexamethasone and the results of previously disclosed sirolimus and dexamethasone. (Suziki, T et al., Stent-based delivery of sirolimus reduces neointimal formation in a porcine coronary model. Circulation. 2001; 104: 1188-1193, Falotico R., U.S. Patent Application 2003/0216699; Table 6.0). This previous study showed no benefit between the combination stent and the single drug eluting stent. Even accompanied by the previously disclosed BX Velocity Bracket data compared to control TriMaxx Significant improvements in stents, in the porcine model with the same overextension ratio, the combination of quetiamus and dexamethasone was substantially superior to the control group and was substantially and statistically superior to the single drug-dissolving stent ZoMaxx ^{T M} .

Example 12 Clinical Experiment

The introduction of stents that deliver a single anti-proliferative agent and subsequent widespread use have reduced the rate of restenosis in the general clinical population by at least 10%. However, there is a clear rationale for the delivery of appropriate drug combinations from the scaffold to treat patients in the general clinical population and to deliver from a large subset of cardiovascular diseases to further reduce restenosis rates and adverse clinical events. For example, it is widely accepted that in patients with diabetes mellitus with stents, the rate of restenosis is significantly increased compared with those without the disease, and the inflammatory response to stenting is present in diabetic and non-diabetic patients (Aggarwal et al. Man, Am. J. Cardiol. 92: 924-929, 2003). In addition, inflammation is associated with acute coronary syndrome (ACS, a term that defines a range of acute myocardial ischemic conditions, including unstable colic, non-ST-segment elevation myocardial infarction, and associated with persistent ST-segment elevation Characteristics of patients with infarction). These patients are often the primary candidates for stent deployment and have significantly higher rates of cyclic ischemia, reinfarction, and subsequent need for repeated PCI procedures relative to the general patient population undergoing percutaneous intervention (PCI). Ultimately, obesity is often associated with pre-inflammatory conditions and endothelial dysfunction. Two conditions are known to be independent predictors of early restenosis after coronary stent placement. In fact, there has been a link between obesity (adipocyte-derived interleukin-6 (IL-6)) and coronary artery disease, indicating an increase in this inflammatory cytokine and CAD development in the patient's subset. Contact (Yudkin et al., Atherosclerosis 148:209-214, 2000).

Diabetes patients are known to exhibit higher levels of inflammatory marker c-reactive protein (CRP) than non-diabetic patients (Aggarwal et al, Dandona and Aljada, Am. J. Cardiol. 90 (Supplement): 27G-33G, 2002). . This protein has been clearly identified as an important inflammatory mediator in patients with coronary artery disease and is a predictor of adverse events in patients with severe unstable colic (Biondi-Zoccai et al., J. Am. Coll. Cardiol. 41: 1071-1077, 2003). CRP is known to stimulate human endothelial cells to produce monocyte chemotactic protein (MCP-1). The release of this mediator is achieved by the injection of monocytes, resulting in a marked inflammatory state, as these cells are intensified and migrate into the endothelial space where they form oxidized low density lipoprotein (LDL). ) foam cells. Plasma IL-6 and tumor necrosis factor-α (TNF-α) are inflammatory cytokines, which are also elevated in obese patients and type 2 diabetic patients. In fact, elevated levels of highly sensitive CRP, IL-6 or serum vascular cell adhesion molecule-1 (VCAM-1) have been associated with increased mortality in patients with coronary artery disease (Roffi and Topol, Eur.Heart J) .25:190-198, 2004). Since neovascular intimal formation (characterized by the restenosis process) has been shown to be exacerbated by inflammation, it is desirable to have a clear utility in diabetic patients using stents that deliver anti-inflammatory drugs to the local vascular environment.

The rupture of atherosclerotic plaque is important for the onset of acute coronary syndrome (Grech and Ramsdale, Br. Med. J., 326: 1259-1260, 2003). Plaque rupture can be induced by increasing the concentration of matrix metalloproteinases secreted by the foam cells, which results in plaque instability and ultimate rupture of the thin fiber cap covering the developmental lesion. In addition, tissue factor activation on the surface of foam cells results in thrombin-forming factor VII. The production of this protein leads to platelet activation and agglutination, as well as the clear formation of fibrinogen into blood fibers and thrombus. The initial concern about the deployment of stents with this configuration does not seem to be found. This is because stent deployment and technical improvements have shown that patients with stents have less ischemic, reinfarction and recurrent angioplasty. Need (Grech and Ramsdale, 2003). The intimate relationship between inflammation and the development of coronary artery injury makes the delivery of anti-inflammatory drugs and anti-proliferative agents an attractive method for treating such patients.

Information on the utility of anti-inflammatory drugs in patients with coronary artery disease has been publicly and clearly described. In the IMPRESS study, event-free survival was significantly enhanced in patients receiving oral treatment with long-term anti-inflammatory drug cortisone (Versaci et al, J. Am. Coll. Cardiol. 40: 1935-1942, 2002). In addition, patients implanted with a stent of the lytic anti-inflammatory drug dexamethasone (Patti et al, Am. J. Cardiol. 95: 502-505, 2005) exhibited a significant reduction in CRP levels within 48 hours of implantation. In CRP value This effect is particularly pronounced and persists in patients with 3 mg/dL.

Scaffolds dissolving dexamethasone have also been shown to have clear benefits in patients with unstable colic (as opposed to stable colic), as reported in the STRIDE trial (Liu et al., 4:265, 2002). As shown by the results of the pig study reported herein, the discovery that the anti-proliferative effect of dexamethasone in vascular muscle smooth muscle cells is combined with the well-known anti-inflammatory effect of this drug indicates the presence of potent anti-proliferative temasis and dexamethasone. The stent is effective in further reducing the rate of restenosis.

Deploying the subgroups in patients with ischemic heart disease diagnosed with stenotic lesions in the coronary arteries and in a subset of clinical populations at higher risk of circulating coronary artery disease and other adverse clinical events support. Other targets for intervention include peripheral vascular disease, including stenosis in the superficial femoral artery, renal artery, ileum, and sub-knee blood vessels. The percutaneous vascular access is used via the femoral or iliac artery to reach the target vessel of the interventional procedure and the introducer cannula is inserted into the vessel. The guidewire is then passed over the target lesion and the balloon catheter is inserted either online or using a rapid exchange system. The surgeon determines the appropriate size of the stent to be implanted by on-line coronary angiography (QCA) or by visual estimation. The stent is deployed using appropriate pressure as indicated by the compliance of the stent, and a post-procedural vascular photograph is then available. When the procedure is completed, the patient is regularly monitored for the presence of a colic condition and any adverse events. The need for repeat procedures is also assessed.

In the Examples, in a model of porcine coronary artery injury with 30% overextension, the antiproliferative activity of the first drug reduced neovascular intimal formation by at least 25% compared to the drug-free stent. In other embodiments, the antiproliferative effect of the first drug complements the antiproliferative effect of the second drug, and in a model of pig injury with 30% overextension, the neovascular intima is formed at least as compared to the drug-free stent. Reduce by 30%.

In an embodiment of the system, the drug has a combination index of less than or equal to 10. In other embodiments, the system further comprises a therapeutic amount of the first drug to the therapeutic amount of the second drug ratio of 1:10 to 10:1 ratio. In other embodiments, the pharmaceutical composition for reducing neovascular intimal hyperplasia is administered topically, and includes quetiamus or everolimus and dexamethasone, wherein quetiamus or everolimus and dexamethasone Rice pine is at a ratio of between about 10:1 and about 1:10. In other embodiments, the pharmaceutical composition for reducing neovascular intimal hyperplasia is administered topically and comprises at least one Moss drug and at least one glucocorticol, and wherein the Moss drug and the glucocorticol are at about 10:1 to about The ratio between 1:10. In addition, endothelialization is accelerated relative to a single drug-eluting stent.

It is to be understood that the foregoing detailed description and the accompanying claims Various changes and modifications to the disclosed embodiments will be apparent to those skilled in the art. Such changes and modifications can be made without departing from the spirit and scope of the invention, including, but not limited to, chemical structures, substituents, derivatives, intermediates, synthetic methods, formulations, and/or methods of using the invention. They have changed and corrected.

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Figure 1 shows the blood concentration ± SEM (n = 3) of a tetrazolium-containing rapamycin analogue taken in monkeys.

2 is a side elevational view showing a stent suitable for use in the present invention. (via BiodivYsio PC coated stent)

Figure 3A is a cross-sectional view of a vascular slice in which a polymer coated stent is placed.

Figure 3B is a cross-sectional view of a blood vessel segment in which a polymeric drug coated stent is placed.

Figure 4 shows a linear blood concentration-time curve for a single elevated intravenous dose of quetiamus in humans.

Figure 5 shows the mean blood concentration-time curve in logarithmic linear scale as the intravenous dose of quetiamus is monotonically elevated in the human body.

Figure 6 shows that the quetiamus Cmax and AUC parameters are proportional to the dose as the intravenous dose in the human body increases monotonically. (Average ABT-578 blood concentration-time curve; log-linear ratio)

Figure 7 shows the mean blood concentration-time curve of quetiamus along with multiple intravenous doses in the human body. (mean on the 14th day ± SD C _max and AUC _0-24 and dose)

Figure 8 shows the average cetamatox blood concentration in the 200, 400, and 800 μg QD (daily) dose groups on Day 1 ( Figure 8a ), Day 14 ( Figure 8b ), and Days 1-14 ( Figure 8c ). - time curve.

Figure 9 shows quetiamus concentration-time data observed on days 1 to 14 for the 800 μg QD dose group. (Average observation and prediction of blood concentration and time curve in accordance with 800μg QD dose group data)

Figures 10 and 11 are graphs showing the effects of quetiamus, dexamethasone, and paclitaxel on LPS-stimulated MCP-1, IL-6, and TNF-α produced by human human monocytes in vitro.

Figure 12 is a graph showing the reduction of the antiproliferative activity of quetiamus in the presence of tacrolimus, which demonstrates that tacrolimus inhibits the antiproliferative activity of quetiamus in vitro in smooth muscle cells.

Figure 13 is a graph showing the anti-proliferative activity of a combination of quetiamus alone, dexamethasone and quetiamus/dexamethasone in human coronary artery smooth muscle cells (hCaSMC; upper panel) and endothelial cells (hCaEC; lower panel). .

Figure 14 shows an isometric effect map of the anti-proliferative activity of the combination of quetiasti/dexamethasone in human coronary artery smooth muscle cells (hCaSMC).

Figure 15 is a graph showing the combined exponential activity of several anti-proliferative activities of a combination of quetiamus/dexamethasone in human coronary artery smooth muscle cells (hCaSMC).

Figures 16 and 17 are graphs of accelerated dissolution rates of stents loaded from quetiamus or dexamethasone. (drug release from self-use quetiamus (10 μg/mm) and dexamethasone (10 μg/mm) loaded stent)

Figure 18 is a graph showing the dissolution rate of quetiamus alone and quetiamus in the presence of dexamethasone. (Seurosol release from a single quetiamus (10 μg/mm) or quetiamus (10 μg/mm) and dexamethasone (10 μg/mm) supported scaffold)

Figure 19 is a bar graph showing the neovascular intimal area (mean ± SEM) 28 days after implantation of the quetiamus/dexamethasone-dissolved stent in pigs compared to the single drug-eluting stent and the control stent.

Figure 20 is a bar graph showing the neovascular intima thickness (mean ± SEM) 28 days after implantation of the quetiamus/dexamethasone-soluble stent in pigs compared to the single drug-eluting stent and the control stent.

Figure 21 is a bar graph showing the percentage of stenotic area (mean ± SEM) 28 days after implantation of quetiamus/dexamethasone-dissolved stent in pigs compared to single drug-eluting stents and control stents.

Figure 22 is a comparison of neovascular intimal area measurements (mean ± SEM; 30%) in two pigs after implantation of quetiamus/dexamethasone-dissolved stent compared to a single drug-eluting stent and control stent. Bar graph with overstretched).

Figure 23 is a comparison of the percentage of stenotic areas (mean ± SEM; 30% overextension) in two pigs after implantation of quetiamus/dexamethasone-dissolved stent compared to a single drug-eluting stent and a control stent. Bar chart.

Figure 24 is a photomicrograph showing the mean response in two pigs after implantation of the quetiamus/dexamethasone-dissolved stent. Figures 24a-e show cross-sectional micrographs of representative blood vessels from pig studies representing the mean neovascular intimal area of each group. FIG 24a: TriMaxx ^TM, stent; 24b: ZoMaxx ^TM stent; 24c: Cypher® stent; 24d: Taxus® holder; 24e: SECRETARY universe Mok: paclitaxel, 10μg / mm: 1μg / mm stent.

Claims (43)

A system for providing controlled release delivery of a medicament for treating or inhibiting neovascular intimal hyperplasia in a vascular lumen, comprising: a composition comprising a therapeutic amount of a first medicament and a therapeutic amount of a second medicament; And wherein the first drug has a therapeutic effect, and the activity of the second drug is supplemented in the presence of the therapeutic amount of the second drug, and wherein the second drug has a therapeutic effect, and the therapeutic amount is The first drug may be supplemented with the activity of the first drug; wherein the therapeutic amount of the first drug and the therapeutic amount of the second drug is a ratio of 1:10 to 10:1; wherein the first drug A zotarolimus; wherein the second drug is dexamathasone. The system of claim 1, wherein the activity of the second drug is anti-inflammatory. The system of claim 1, wherein the activity of the first drug is anti-proliferative. The system of claim 1 wherein the composition is associated with a medical device. The system of claim 4, wherein the medical device comprises a stent. The system of claim 4, wherein the medical device comprises an angioplasty balloon. The system of claim 5, wherein the stent further comprises a coating on at least a portion of the surface of the stent. The system of claim 7, wherein the composition is associated with the coating. The system of claim 7, wherein the coating comprises a polymer. The system of claim 9, wherein the polymer comprises a phosphonium choline polymer. The system of claim 9, wherein the polymer is selected from the group consisting of fluoropolymers, polyacrylates, polyoxyxides, resins, nylons, and poly(decylamines). The system of claim 5, wherein the therapeutic amount of zotarolimus is at least 1 μg per millimeter of stent. The system of claim 5, wherein the therapeutic amount of the second medicament is at least 0.5 μg per millimeter of stent. The system of claim 1 further comprising a third therapeutic drug or substance. The system of claim 14, wherein the third therapeutic agent is selected from the group consisting of an anti-proliferative agent, an anti-platelet agent, an anti-inflammatory agent, an anti-lipemic agent, an antithrombotic agent, a thrombolytic agent, a salt thereof, or Any combination of them. The system of claim 14, wherein the third therapeutic agent is a glucocorticol comprising a group consisting of: methylprednisolone, prednisolone, prednisone, triamcinolone (triamcinolone), dexamethasone, mometasone, beclomethasone, ciclesonide, bedesonide, triamcinolone, clobeta Dobetasol, flunisolide, loteprednol, budesonide (budesonide), fluticasone, a salt thereof, or any combination thereof. The system of claim 14, wherein the third therapeutic agent is a steroid hormone comprising estradiol and a salt thereof, or any combination thereof. The system of claim 14, wherein the third therapeutic agent is a member of a group consisting of small molecules and biological agents that reduce inflammatory cytokine activity. The system of claim 14, wherein the third therapeutic agent comprises an anti-TNFα therapeutic agent consisting of a group of adalimumab, an anti-MCP-1 therapeutic agent, a CCR2 receptor antagonist, and an anti-TNFa therapeutic agent. An IL-18 therapeutic, an anti-IL-1 therapeutic, and a salt thereof, or any combination thereof. The system of claim 14, wherein the third therapeutic agent comprises an anti-proliferative agent selected from the group consisting of cyclophosphamide, chlorambucil, busulfan, and cardio Carmustine, lomustine, methotrexate, fluorouracil, cytarabine, thioglycol, pentostatin, vinblastine, vincristine , doxorubicin, bleomycin, mitomycin, cisplatin, procarbazine, etoposide, teniposide; Salt and any combination thereof. The system of claim 14, wherein the third therapeutic agent comprises an antiplatelet agent selected from the group consisting of abciximab, eptifibatide, tirofiban Dipyridamole Dipyridamole, clopidogrel, ticlopidine, acetyl sulphate, cilostazol; salts thereof, and any combination thereof. The system of claim 14, wherein the third therapeutic agent comprises an anti-inflammatory agent selected from the group consisting of: dexamethasone, hydrocortisone, fluticasone, clobetasol, mometasone, Estradiol, acetaminophen phenol, ibuprofen, naproxen, sulindac, piroxicam, mefanamic acid; IL-1, IL - 2, antibodies to IL-8, IL-15, IL-18 and TNF; salts thereof and any combination thereof. The system of claim 14, wherein the third therapeutic agent comprises an antithrombotic agent selected from the group consisting of unsegmented heparin, low molecular weight heparin, clivarin, dalteparin ), enoxaparin, nadroparin, tinzaparin, argatroban, hirudin, hirulog, hirugen; salts thereof Any combination of them. The system of claim 14, wherein the third therapeutic agent comprises an anti-lipemic agent selected from the group consisting of: mevastatin, lovastatin, simvastatin, pravud Pravastatin, fluvastatin, fenofibrate, clofibrate, gemfibrozil, nicotinic acid, probucol; salts thereof and any combination. The system of claim 14, wherein the third therapeutic agent comprises a selected one a thrombolytic agent consisting of a group consisting of streptokinase, urokinase, prourokinase, alteplase, reteplase, tenectalpase, salts thereof, and any combination. The system of claim 5, wherein the therapeutic amount of the second medicament is at least 1 μg per millimeter of stent. The system of claim 3, wherein the antiproliferative activity of the first drug reduces neovascular intimal formation by at least 25% in a model of porcine coronary artery injury having a 30% overextension compared to a drug-free stent. . The system of claim 1, wherein the drug has a combination index of less than or equal to 10. The system of claim 3, wherein the anti-proliferative effect of the first drug complements the anti-proliferative effect of the second drug, and may be regenerated in a pig injury model with 30% overextension compared to a drug-free stent Endocardial formation is reduced by at least 30%. A pharmaceutical composition for topical administration to reduce neovascular intimal hyperplasia comprising quetiamus and dexamethasone, wherein the quetiamus and the dexamethasone are between about 10:1 and about 1 : 10 ratio. The pharmaceutical composition of claim 30, when the pharmaceutical composition is delivered by a drug-eluting stent, accelerates endothelialization compared to a single drug. The pharmaceutical composition of claim 30, further comprising a third therapeutic drug or substance. The pharmaceutical composition of claim 32, wherein the third therapeutic agent is selected from the group consisting of an anti-proliferative agent, an anti-platelet agent, an anti-inflammatory agent, an anti-lipemic agent, an antithrombotic agent, a thrombolytic agent, Salt or its Any combination. The pharmaceutical composition of claim 32, wherein the third therapeutic agent is a glucocorticol comprising methylprednisolone, prednisolone, prednisone, triamcinolone, dexamethasone, mometasone, and beclomethasone , ciclesonide, bettydine, triamcinolone, clobetasol, flunisolide, loteprednol, budesonide, fluticasone, a salt thereof, or any combination thereof. The pharmaceutical composition of claim 32, wherein the third therapeutic agent is a steroid hormone comprising estradiol and salts, prodrugs and derivatives thereof, or any combination thereof. The pharmaceutical composition of claim 32, wherein the third therapeutic agent is a member of a group of small molecules and biological agents that reduce inflammatory cytokine activity. The pharmaceutical composition according to claim 32, wherein the third therapeutic agent comprises an anti-TNFα therapeutic agent comprising adalimumab, an anti-MCP-1 therapeutic agent, a CCR2 receptor antagonist, an anti-IL-18 therapeutic agent, and an anti-IL - 1 therapeutic agent and its salt or any combination thereof. The pharmaceutical composition of claim 32, wherein the third therapeutic agent comprises an anti-proliferative agent selected from the group consisting of cyclophosphamide, chlorambucil, busulfan, carmust, and lomo Statin methylamine, fluorouracil, cytarabine, thioglycol, pentastatin, vinblastine, vincristine, doxorubicin, bleomycin, mitomycin, cisplatin, procarbazine, relying on Bovine glycosides, teniposide; salts thereof and any combination thereof. The pharmaceutical composition of claim 32, wherein the third therapeutic agent comprises an antiplatelet agent selected from the group consisting of abciximab, ezetidine Non-bapeptide, tirofiban, dipyridamole, clopidogrel, ticlopidine, acetyl sulphate, cilostazol; salts thereof or any combination thereof. The pharmaceutical composition of claim 32, wherein the third therapeutic agent comprises an anti-inflammatory agent selected from the group consisting of dexamethasone, hydrocortisone, fluticasone, clobetasol, mometasone, and estrogen Glycol, acetaminophen phenol, ibuprofen, naproxen, sulindac, piroxicam, mefenamic acid; IL-1, IL-2, IL-8, IL-15, IL-18 and TNF An antibody; a salt thereof and any combination thereof. The pharmaceutical composition of claim 32, wherein the third therapeutic agent comprises an anti-thrombotic agent selected from the group consisting of unsegmented heparin, low molecular weight heparin, clover heparin, dalteparin, enoxa Heparin, nadroparin, tinzaparin, argatroban, hirudin, black lulu, black root; its salts and any combination thereof. The pharmaceutical composition of claim 32, wherein the third therapeutic agent comprises an anti-lipemic agent selected from the group consisting of: mevastatin, lovastatin, simvastatin, pravastatin, fluvastatin, non- Nobel, clofibrate, gemfibrozil, nicotinic acid, probucol; its salts and any combination thereof. The pharmaceutical composition of claim 32, wherein the third therapeutic agent comprises a thrombolytic agent selected from the group consisting of: streptokinase, urokinase, prourokinase, alteplase, reteplase, Naproxase, its salts, and any combination thereof.