HK1056161B - Amidino compound and salts thereof useful as nitric oxide synthase inhibitors - Google Patents

Description

Amidino compounds and salts thereof useful as nitric oxide synthase inhibitors

This application claims priority from U.S. provisional patent application No. 60/191,923, filed on 24/3/2000, which is incorporated herein by reference.

Background

Technical Field

The present invention relates to amidino compounds useful as nitric oxide synthase inhibitors.

Prior Art

Since the early 80 s of the 20 th century, it was known that acetylcholine-induced vasodilation was dependent on the vascular endothelium. Endothelial cell vasopressin (EDRF), now known as Nitric Oxide (NO), is produced in the vascular endothelium by Nitric Oxide Synthase (NOs). The activity of NO as a vasodilator has been known for over 100 years. In addition, NO is an active substance (species) derived from isoamyl nitrite, nitroglycerin and other nitrovasodilators. The identification of EDRF as NO is consistent with the discovery of the biochemical pathway through which NO is synthesized from the amino acid L-arginine using the enzyme NO synthase.

Nitric oxide is an endogenous stimulator of soluble guanylate cyclase. In addition to endothelium-dependent relaxation, NO is involved in a number of biological actions, including phagocytic cytotoxicity and cell-to-cell communication within the central nervous system.

There are at least three types of NO synthases:

(i) endothelial constitutive Ca⁺⁺Calmodulin-dependent enzymes, which release NO in response to a receptor or physical stimulus.

(ii) Constitutive Ca localized in the brain⁺⁺Calmodulin-dependent enzymes, which release NO in response to a receptor or physical stimulus.

(iii) Ca-independence induced after endotoxin and cytokine activation of vascular smooth muscle, macrophages, endothelial cells and a large number of other cells⁺⁺Once expressed, this inducible nitric oxide synthase (hereinafter "iNOS") produces NO continuously over a long period of time.

The NO released by each of the two constituent enzymes acts as a transduction mechanism that undergoes several physiological responses. NO produced by the inducible enzyme is a cytotoxic molecule to tumor cells and invading microorganisms. It has also been shown that the side effects of excessive NO production, especially pathological vasodilation and tissue damage, can be caused in large part by NO synthesized by iNOS.

There is increasing evidence that NO may be involved in cartilage degeneration caused by certain diseases such as arthritis, and it is also known that NO synthase is increased in rheumatoid arthritis and osteoarthritis.

Some of the proposed NO synthase inhibitors for use in therapy are non-selective; they inhibit constitutive and inducible NO synthase. The use of such non-selective NO synthase inhibitors requires great care to avoid the potentially serious consequences of over-inhibition of constitutive NO synthase, including hypertension and possible thrombosis and tissue damage. Particularly in the treatment of toxic shock with L-NMMA, it is recommended that the patient must be monitored continuously for blood pressure throughout the course of treatment. Thus, the therapeutic utility of non-selective NO synthase inhibitors is premised on the assumption that, with appropriate precautions, NO synthase inhibitors are selective in the sense that their inhibition of inducible NO synthase is much greater than the inhibition of the constitutive NO synthase isoforms, and thus have even greater therapeutic benefit and are easier to use (S.Moncada and Higgs, E.FASEB J., 9, 1319-.

Each of the following publications discloses compounds that inhibit nitric oxide synthesis and preferentially inhibit the isoform of inducible nitric oxide synthase:

PCT patent application No. WO96/35677,

PCT patent application No. WO96/33175,

PCT patent application No. WO96/15120,

PCT patent application No. WO95/11014,

PCT patent application No. WO95/11231,

PCT patent application No. WO99/46240,

PCT patent application No. WO95/24382,

PCT patent application No. WO94/12165,

PCT patent application No. WO94/14780,

PCT patent application No. WO93/13055,

PCT patent application No. WO99/62875,

European patent No. EP0446699A1,

U.S. Pat. No. 5,132,453,

U.S. Pat. No. 5,684,008,

U.S. Pat. No. 5,830,917,

U.S. Pat. No. 5,854,251,

U.S. Pat. No. 5,863,931,

U.S. Pat. No. 5,919,787,

U.S. Pat. No. 5,945,408,

U.S. patent No. 5,981,511.

PCT patent application No. WO95/25717 discloses certain amidino derivatives useful for the inhibition of inducible nitric oxide synthase.

PCT patent application No. WO99/62875 discloses other amidino derivatives useful for the inhibition of inducible nitric oxide synthase.

Summary of The Invention

Compounds that have the advantage of being very effective as iNOS inhibitors have been found in human cartilage explant assays, a model for osteoarthritis. At the same time, the compounds of the invention surprisingly do not penetrate certain non-target organs in the test system, especially when compared to the compounds of WO 95/25717. The surprising difference in the expected selection between the target organ (cartilage) and other organs is an unexpected advantage of the compounds of the present invention.

In a broad aspect, the present invention is directed to novel compounds, pharmaceutical compositions and methods of inhibiting or modulating nitric oxide synthesis using the compounds and compositions, which comprise administering to a patient in need of such inhibition or modulation a compound that preferentially inhibits or modulates an isoform of inducible nitric oxide synthase over an isoform of constitutive nitric oxide synthase. It is another object of the present invention to reduce nitric oxide levels in a patient in need of such reduction. The compounds of the present invention have useful nitric oxide synthase inhibitory activity and are expected to be useful in the treatment or prevention of diseases or conditions in which nitric oxide is synthesized or over-synthesized as part of the etiology.

In one embodiment, the present invention provides a compound, or salt thereof, having a structure corresponding to formula 1:

wherein:

x is selected from the group consisting of-S-, -S (O) -and-S (O)₂-；

R²Is selected from C₁-C₆Alkyl radical, C₂-C₆Alkenyl radical, C₂-C₆Alkynyl, C₁-C₅alkoxy-C₁Alkyl and C₁-C₅alkylthio-C₁An alkyl group;

as for R³And R⁸：

R⁸Is selected from-OR¹⁴and-N (R)¹⁵)(R¹⁶) (ii) a And R is³Selected from the group consisting of-H, -OH, -C (O) -R¹⁷、-C(O)-O-R¹⁸and-C (O) -S-R¹⁹(ii) a Or

R⁸is-N (R)²⁰) -and R³is-C (O) -, wherein R⁸And R³Form a ring together with the atoms to which they are attached; or

R⁸is-O-and R³is-C (R²¹)(R²²) -, wherein R⁸And R³Form a ring together with the atoms to which they are attached;

if R is³is-C (R)²¹)(R²²) -, then R⁴is-C (O) -O-R²³(ii) a Otherwise R⁴is-H;

R¹、R⁵、R⁶and R⁷Independently selected from-H, halogen, C₁-C₆Alkyl radical, C₂-C₆Alkenyl radical, C₂-C₆Alkynyl and C₁-C₅alkoxy-C₁An alkyl group;

R⁹and R¹⁰Independently selected from-H, C₁-C₆Alkyl radical, C₂-C₆Alkenyl radical, C₂-C₆Alkynyl and C₁-C₅alkoxy-C₁An alkyl group;

as for R¹¹And R¹²：

R¹¹Selected from the group consisting of-H, -OH, -C (O) -O-R²⁴and-C (O) -S-R²⁵(ii) a And R is¹²Selected from the group consisting of-H, -OH, -C (O) -O-R²⁶and-C (O) -S-R²⁷(ii) a Or

R¹¹is-O-and R¹²is-C (O) -, wherein R¹¹And R¹²Form a ring together with the atoms to which they are attached; or

R¹¹is-C (O) -, and R¹²is-O-wherein R¹¹And R¹²Form a ring together with the atoms to which they are attached; and

R¹³is C₁An alkyl group;

R¹⁴is selected from-H and C₁-C₆An alkyl group; wherein when R is¹⁴Is C₁-C₆When alkyl, R¹⁴Optionally substituted with one or more groups selected from cycloalkyl, heterocyclyl, aryl and heteroaryl;

as for R¹⁵And R¹⁶：

R¹⁵Selected from-H, alkyl and alkoxy; and R is¹⁶Selected from the group consisting of-H, -OH, alkyl, alkoxy, -C (O) -R^27a、-C(O)-O-R²⁸and-C (O) -S-R²⁹(ii) a Wherein when R is¹⁵And R¹⁶When independently alkyl or alkoxy, R¹⁵And R¹⁶Independently optionally substituted with one or more groups thereof selected from cycloalkyl, heterocyclyl, aryl and heteroaryl; or

R¹⁵is-H; and R is¹⁶Selected from the group consisting of cycloalkyl, heterocyclyl, aryl and heteroaryl;

R¹⁷、R¹⁸、R¹⁹、R²⁰、R²¹、R²²、R²³、R²⁴、R²⁵、R²⁶、R²⁷、R^27a、R²⁸and R²⁹Independently selected from-H and alkyl, said alkyl being optionally substituted with one or more groups selected from cycloalkyl, heterocyclyl, aryl and heteroaryl; and is

When R is¹、R²、R³、R⁴、R⁵、R⁶、R⁷、R⁸、R⁹、R¹⁰、R¹¹、R¹²、R¹³、R¹⁴、R¹⁵、R¹⁶、R¹⁷、R¹⁸、R¹⁹、R²⁰、R²¹、R²²、R²³、R²⁴、R²⁵、R²⁶、R²⁷、R^27a、R²⁸And R²⁹When any of (a) is independently a group selected from alkyl, alkenyl, alkynyl, alkoxy, alkylthio, cycloalkyl, heterocyclyl, aryl and heteroaryl, the group is optionally substituted with one or more substituents selected from-OH, alkoxy and halogen.

Another embodiment provides a method of treating or preventing a disease associated with inflammation, said method comprising treating a patient in need thereof with a therapeutic or prophylactic amount of a compound of the invention for a disease associated with inflammation.

In another embodiment, the invention provides a compound of formula (I)1A method of preparing a compound, wherein the method comprises administering a compound having a formula corresponding to23Alkyl acetimidate of structure (alkyl acetimidate):

wherein R is³¹Is C₁-C₆Alkyl radical, treatment having a formula22A diamine compound of the structure of (a):

. If desired, the treatment can be carried out in the presence of an acid or a base, preferably in the presence of a base.

In another embodiment of the present invention, there is provided a process for preparing a compound having a formula corresponding to22A method of producing a diamine compound of the structure (1) or a salt thereof:

wherein R is³⁰Selected from the group consisting of-H, -OH, -C (O) -R¹⁷、-C(O)-O-R¹⁸and-C (O) -S-R¹⁹And the other substituents are as defined above, wherein the process comprises treating a compound having a structure corresponding to formula24A protected diamine compound of the structure:

wherein R is³³Selected from-H and protected amino; r³²Is a protected amino group; and R¹⁴Is selected from-H and C₁-C₆An alkyl group; wherein when R is¹⁴Is C₁-C₆When alkyl, R¹⁴Optionally substituted with one or more groups selected from cycloalkyl, heterocyclyl, aryl and heteroaryl; wherein the treatment is with a deprotecting reagent, thereby producing the diamine compound.

Detailed Description

Formula (I) among others1The compounds will be useful in the treatment of inflammation in a patient or in the treatment of other nitric oxide synthase-induced disorders, e.g., as analgesics for pain and headache, or as antipyretics for fever. For example, the compounds of the present invention will be used to treat arthritis, including but not limited to rheumatoid arthritis, spondyloarthropathies, gouty arthritis, osteoarthritis, systemic lupus erythematosus, juvenile arthritis, acute rheumatoid arthritis, enteropathic arthritis, neuropathic arthritis, psoriatic arthritis, and suppurative arthritis. Conditions in which compounds of the invention would provide advantages in inhibiting the production of NO from L-arginine include arthritic conditions.

The compounds of the invention will further be useful in the treatment of asthma, bronchitis, menstrual cramps (e.g., dysmenorrhea), premature labor, tendonitis, bursitis, skin-related conditions such as psoriasis, eczema, burns, sunburn, dermatitis, pancreatitis, hepatitis and post-operative inflammation including that following ophthalmic surgery such as cataract surgery and refractive surgery. The compounds of the invention are also useful in the treatment of gastrointestinal disorders such as inflammatory bowel disease, crohn's disease, gastritis, irritable bowel syndrome and ulcerative colitis. The compounds of the present invention will be useful for the prevention or treatment of cancers such as colorectal cancer, breast cancer, lung cancer, prostate cancer, bladder cancer, neck cancer and skin cancer. The compounds of the present invention will be useful in the treatment of inflammation and tissue injury, such as vascular disease, migraine, adventitial inflammation of nodal artery, thyroiditis, aplastic anemia, hodgkin's disease, sclerodoma, rheumatic fever, type I diabetes, neuromuscular junction disease (meniscularjunction) including myasthenia gravis, white matter disorders including multiple sclerosis, sarcoidosis, nephrotic syndrome, herbecitt's syndrome, polymyositis, gingivitis, nephritis, allergy, post-injury swelling, myocardial ischemia, and the like. The compounds of the present invention are also useful in the treatment of ocular diseases such as glaucoma, retinitis, retinopathy, uveitis, ocular photophobia, and inflammation and pain associated with acute injury to ocular tissues. Of particular importance for the use of the compounds of the present invention is the treatment of glaucoma, especially the symptoms of glaucoma caused by nitric oxide production, such as nitric oxide mediated nerve damage. The compounds of the invention are also useful in the treatment of pneumonia, such as that associated with viral infections and cystic fibrosis. The compounds of the present invention are also useful in the treatment of certain central nervous system disorders, such as cortical dementias (cortical dementias) including Alzheimer's disease and central nervous system injuries caused by stroke, ischemia and trauma. The compounds of the present invention are useful as anti-inflammatory agents, such as for the treatment of arthritis, with the added benefit of minimal deleterious side effects. These compounds are also useful in the treatment of allergic rhinitis, respiratory distress syndrome, endotoxic shock syndrome and atherosclerosis. The compounds are also useful for treating pain, but not limited to, postoperative pain, dental pain, muscle pain, and pain caused by cancer. The compounds will be useful in the prevention of dementias such as Alzheimer's disease.

In addition to use as human therapeutics, these compounds are also useful in veterinary therapeutics in domestic companion animals, exotic animals and farm animals, including mammals, rodents, and the like. More preferred animals include horses, dogs, and cats.

The compounds of the invention may also be used, for example, in combination with steroids, NSAIDs, COX-2 selective inhibitors, 5-lipoxygenase inhibitors, LTB₄Antagonists and LTA₄Hydrolase inhibitors are used together in combination therapy, partially or completely replacing other conventional anti-inflammatory therapies.

Other conditions in which compounds of the invention will provide the advantage of inhibiting the inhibition of NO include cardiovascular ischemia, diabetes (type I or type II), congestive heart failure, myocarditis, atherosclerosis, migraine, glaucoma, aortic aneurysm, reflux esophagitis, diarrhoea, irritable bowel syndrome, cystic fibrosis, emphysema, asthma, bronchiectasis, hyperalgesia (allodynia), cerebral ischemia (bifocal ischemia, thrombotic stroke and global ischemia (e.g. secondary cardiac arrest), multiple sclerosis and other diseases of the central nervous system caused by NO, such as Parkinson's disease Hyperbaric oxygen convulsions and intoxications, dementias such as Alzheimer's disease and AIDS-related dementia, cachexia, West German chorea, Huntington's chorea, amyotrophic lateral sclerosis, Korsakoff's disease, dementia associated with cerebrovascular disease, sleep disorders, schizophrenia, depression or other symptoms associated with premenstrual syndrome (PMS), anxiety and septic shock.

The compounds of the invention are also useful in the treatment of pain including somatogenic (nociceptive or neuropathic) pain, including acute and chronic pain. Nitric oxide inhibitors may be used for any condition including neuropathic pain that is traditionally administered with a common NSAID or opioid analgesic.

Other diseases or conditions which may be advantageously treated with the compounds of the present invention also include opiate tolerance in patients in chronic need of opioid analgesics and benzodiazepine * tolerance in patients with benzodiazepine * class, and the treatment or prevention of other addictive behaviors such as nicotine addiction, alcoholism and eating disorders. The compounds and methods of the invention are also useful for the treatment or prevention of drug withdrawal syndromes, such as the withdrawal symptoms of opiate, alcohol or tobacco addiction. The compounds of the present invention are also useful for preventing tissue damage when treated in combination with antibacterial or antiviral agents.

The compounds of the invention are also useful for inhibiting the production of NO from L-arginine, including systemic hypotension associated with septic and/or toxic hemorrhagic shock induced by a variety of agents; treatment with cytokines such as TNF, IL-1 and IL-2; and in transplantation therapy, as an adjuvant to short-term immunosuppression.

The invention also relates to the use of the compounds of the invention for the treatment and prevention of neoplasia. Neoplasias that may be treated or prevented with the compounds and methods of the present invention include brain cancer, bone cancer, leukemia, lymphoma, epithelial cell-derived neoplasias (epithelial cancers) such as basal cell carcinoma, adenocarcinoma, gastrointestinal cancers such as lip cancer, oral cancer, esophageal cancer, small bowel cancer and stomach cancer, colon cancer, liver cancer, bladder cancer, pancreatic cancer, ovarian cancer, cervical cancer, lung cancer, breast cancer, and skin cancers such as squamous cell and basal cell carcinoma, prostate cancer, renal cell carcinoma, and other cancers known to affect epithelial cells throughout the body. Preferred neoplasias are selected from gastrointestinal cancer, liver cancer, bladder cancer, pancreatic cancer, ovarian cancer, prostate cancer, cervical cancer, lung cancer, breast cancer and skin cancer, such as squamous cell and basal cell carcinoma. The compounds and methods of the present invention are also useful for treating fibrosis caused by radiation therapy. The compounds and methods of the invention are useful for treating patients with adenomatous polyps, including patients with Familial Adenomatous Polyposis (FAP). In addition, the compounds and methods of the invention are useful for preventing polyps from forming in patients at risk for FAP.

The combination therapy of the compounds of the present invention with another antineoplastic agent will produce a synergistic effect or reduce the toxic side effects associated with chemotherapy by reducing the therapeutic dose of the drug required for efficacy to cause the side effects, or by directly reducing the symptoms of toxic side effects produced by the drug causing the side effects.

The compounds of the present invention may also be used as an adjuvant to radiotherapy to reduce side effects or increase effectiveness. In the present invention, another agent that may be combined with a compound of the present invention includes any therapeutic agent that inhibits the enzyme cyclooxygenase-2 ("COX-2"). Preferably, such COX-2 inhibitors selectively inhibit COX-2 over the enzyme cyclooxygenase-1 ("COX-1"). Such COX-2 inhibitors are referred to as "COX-2 selective inhibitors". More preferably, in an in vitro assay, the compounds of the invention may be used in combination therapy with a COX-2 selective inhibitor, wherein the COX-2 selective inhibitor selectively inhibits COX-2 at a ratio of at least 10: 1, more preferably at least 30: 1, even more preferably at least 50: 1 relative to inhibition of COX-1. COX-2 selective inhibitors for use in combination therapy with the compounds of the invention include celecoxib, valdecoxib, deracoxib, etoricoxib, rofecoxib, ABT-963(2- (3, 4-difluorophenyl) -4- (3-hydroxy-3-methyl-1-butoxy) -5- [4- (methylsulfonyl) phenyl-3 (2H) -pyridazinone; described in PCT patent application No. WO 00/24719), or meloxicam. The compounds of the invention may also be advantageously used in combination therapy with COX-2 selective inhibitors such as prodrugs of parecoxib.

For example, another chemotherapeutic agent that can be used in combination with a compound of the invention can be selected from the following table, which is not all inclusive and is non-limiting:

alpha-Difluoromethylornithine (DFMO), 5-FU-fibrinogen, acanthifolinic acid, aminothiadiazole, brequinar sodium, carmofur, Ciba-Geigy CGP-30694, cyclopentylcytosine, cytarabine phosphate, cytarabine arabinoside conjugate, Lilly DATHF, Merrel Dow DDFC, dezaguanine, dideoxycytidine, dideoxyguanosine, didox, Yoshitomi DMDC, deoxyfluorouridine, Wellcome EHNA, Merck & Co.EX-015, fazabine, floxuridine, fludarabine phosphate, 5-fluorouracil, N- (2' -furyl (furanidyl) -5-fluorouracil), Daiichi Seiyaku FO-152, isopropylpyrrazine, Lilly LY-188011, Lilly-264618, methoprim, methotrexate, amizapine, Weildini, NCNSI-612567, NCNSI-366754, NCNSI-363657, NCC-39661, NCC 39661, NCI-39661, Warner-LambertPALA, pentostatin, pirtroxin, plicamycin, Asahi Chemical PL-AC, TakedaTAC-788, thioguanine, thiazolulin, Erbamont TIF, tritrexate, tyrosine kinase inhibitors, tyrosinase inhibitors, Taiho UFT, uricetin, Shionogi254-s, oxo-phosphoramide analogs, altretamine, anastroketone, Boehringer Mannheim BBR-2207, bestreucil, Brodotitanium, Wakunaga CA-102, carboplatin, nitrosourea nitrogen mustard, Chinoin-139, Chinoin-153, chlorambucil, cyclophosphamide, American Cyanamid CL-286558, Sanofi CY-233, cisplatin, Degussa D-19-384, Sudomoto DAthion, Diphenylamine 2 (Diphenylspirone), Diphenbutamine A-21152, Erosamine derivatives, Ergonomic acid, platinum-D-09, and platinum-D-09, Estramustine sodium phosphate, fotemustine, Unimed G-6-M, ChinoinGYKI-17230, hepsul-fam, ifosfamide, iproplatin, lomustine, macsfamide, dibromodulcitol, Nippon Kayaku NK-121, NCI NSC-264395, NCINSC-342215, oxaliplatin, Upjohn PCNU, melphalan, Proter PTT-119, ramustine, semustine, SmithKline SK & F-101772, Yakult Honsha SN-22, spiromustine, Tanabe Seiyaku TA-077, Tatemustine, temozolomide, temozolone, teplatin, trimelamol, Taiho 4181-A, aclarubicin, actinomycin D, actinoplanone, Erbamemont ADemont R-456, atropine AN derivatives, Ajintan-6-M, ChinoinGYKI-17230, rapamycin-685-201, Myrothecin-685-A, and Amomum japonicumin, Bristol-Myers BMY-25067, Bristol-Myers BMY-25551, Bristol-Myers BMY-26605, Bristol-Myers BMY-27557, Bristol-Myers BMY-28438, bleomycin sulfate, patatin-1, Taiho C-1027, calicheamicin, chromomycin, actinomycin D, daunorubicin, Kyowa Hakko DC-102, Kyowa Hakko DC-79, Kyowa Hakko DC-88A, Kyowa Hakko DC89-A1, Kyowa Hakko DC92-B, ditristaruubicin B, Shiyogi DOB-41, doxorubicin-fibrinogen, elsamicin-A, epirubicin, amierb, epothilone A21954, furcesackiecin-359007, gentamycin FK-5631, gentamycin FR-5631, gentamycin FR-35977, gentamycin FR-5635, gentamycin, Cryptobrecin, kazusamycin, kesarhrodins, Kyowa Hakko KM-5539, Kirin BreweryKRN-8602, Kyowa Hakko KT-5432, Kyowa Hakko KT-5594, Kyowa Hakko KT-6149, American Cyanamid LL-D49194, Meiji SeikaME2303, minoritol, mitomycin, mitoxantrone, SmithKline M-TAG, neoenactin, Japanese Kayaku NK-313, Japanese Kayaku NKT-01, SRIInteronal NSC-357704, lysin, oxomycin, peplomycin, pilatin, birubicin, podhramycin, pyradamycin A, Pharmamycin RA-I, rapamycin, polyoxin, paromycin-B, Taiho 4181, paromycin-S-D-, Takeda TAN-868A, terpentectin, thrazine, triclozarin A, Upjohn U-73975, Kyowa Hakko UCN-10028A, Fujisawa WF-3405, YoshtomimiY-25024 zorubicin, alpha-carotene, alpha-difluoromethyl-arginine, Avermen A, BiotecAD-5, Kyorin AHC-52, thermolysin, amonafide, amphetanilile, amsacrine, Angiostat, anomycin, antineoplastin, antineoplaston A10, antineoplaston A2, antineoplaston A3, antineoplaston A5, antineoplaston AS2-1, Henkelprolide, alfasin glycin glycinate, asparaginase, Avalar Carcassin, Britropha, Bripentulosin BW, Brillotre, Melothrix Gmbycine-2050, Melothrix-2303, Melothrix-230x, Melothrix-351, Melothrix-1 e, Melothrix, Warner-Lambert CI-937, Warner-Lambert CI-941, Warner-Lambert CI-958, kraflururon, claviridone, ICN 1259, ICN 4711, Contracan, Yakult Honsha CPT-11, crisnatol, curaderm, cytochalasin B, cytarabine, cytocytin, Merz D-609, DABIS maleate, dacarbazine, daptomazine, didymin-B, hematoporphyrin ether (dihematoporphin ether), dihydrodelrin, dinalin, distamycin, topharmar DM-341, Toyo Pharmar DM-75, Daiichi Seiyaku DN-9693, ellilobin, acetic acid, glucurochrome HO, Fuyograph HO-N3543, Hodgrain-N63178, Glycine phosphate, Glycine A-857, Glycine A-III, Glycine A-3543, Glycine A-III, Glycine A-III, Glycine-III, N-III, N-III-I-III, N-III-IV, N-III-2-III, Hydroxyurea, BTG ICRF-187, isomofoxin, isoglutamic acid, isotretinoin, Otsuka JI-36, Ramot K-477, OtsuakK-76COONa, Kureha Chemical K-AM, MECT Corp KI-8110, American cyanamid L-623, leupeptin, lonidamine, Lundbeck LU-23-112, Lilly LY-186641, NCI (US) MAP, marycin, Merrel Dow MDL-27048, Medmeco DR-340, merbarone, merocyanine derivatives, methylanilinoacridine, Molecular Genetics MGI-136, minicivin, mitonafide, mitoquinolone, mopidanol, Movea amine, Zenyaku Koyo MST-16, N- (Nissxanthoyl) amino acids, Milzhi-021, N-malonyl alanine-190, N-alanine-arginine derivatives, Tatsukazak JI L-36, clonidine, Lu-L-D-L-D-L-H-L, NCI NSC-145813, NCINSC-361456, NCI NSC-604782, NCI NSC-95580, octreotide, OnoONO-112, oquinolocine, Akzo Org-10172, pancratistatin, pazeliptin, Warner-LambertPD-111707, Warner-LambertPD-115934, Warner-LambertPD-131141, Pierre Fabre PE-1001, ICRT peptide D, piroctone, polyhedrin, polyperacid, Efamol porphyrin, probimane, procarbazine, proglumide, Invitron ANC protease connexin I, Tobishi RA-700, Razoxan, Sappoweries, RBS, Restricin-P, retelliptine, retinoic acid, Rhosenc-Poulc-49RP-4925, Klinelsberg-94, Klaus-S-10094, S-94, S-K-S-94, S-S, Hemostatic alcohol dione (strypoldinone), hemostatic alcohol dione (Stypoldione), Suntory SUN 0237, Suntory SUN 2071, superoxide dismutase, Toyama T-506, Toyama T-680, taxol, Teijin TET-0303, teniposide, thalistine, Eastman Kyowa T-29, tocotrienol, Toponin, Teijin TT-82, Kyowa Hakko UCN-01, Kyowa Hakko UCN-1028, ukrain, Eastman Kodak USB-006, vinblastine sulfate, vincristine, vindesine, vinestramide, vinorelbine, vintriptol, vinzolidine, withanolides, Yamanouchi YM-534, urogounylin, combretastatin, dolastatin, demethoxydaunorubicin, epidaunorubicin, estramustine, cyclophosphamide, 9-amino-2- (S) -camptothecin, topotecan, irinotecan (Camptosar), exemestane, 6-D-tryptorelin (trptorelin), or an omega-3 fatty acid.

Examples of radioprotective agents that may be used in combination therapy with the compounds of the present invention include AD-5, adchnon, amifostine analogs, detox, dimesna, l-102, MM-159, N-acylated-dehydroalanine, TGF-Genentech, thiabendamod, amifostine, WR-151327, FUT-187, percutaneous ketoprofen, nabumetone, superoxide dismutase (Chiron), and superoxide dismutase Enzon.

The compounds of the invention are also useful in the treatment or prevention of diseases or disorders associated with angiogenesis, such as, for example, tumor growth, metastasis, macular degeneration, and atherosclerosis.

In another embodiment, the invention also provides combination therapies for treating or preventing an ocular disease or condition, such as glaucoma. For example, the compounds of the present invention will be advantageously used in combination therapy with a drug that reduces intraocular pressure in patients suffering from glaucoma. Such intraocular pressure-reducing agents include, but are not limited to, latanoprost, travoprost, bimatoprost, or unoprost. The combination therapy of the compounds of the present invention plus an intraocular pressure-lowering agent will be effective because each is believed to exert its effect through a different mechanism.

In another combination therapy of the invention, the compounds of the invention may be used in combination therapy with antihyperlipidemic or cholesterol lowering drugs such as benzothiepine or benzothiazepine * (benzothiazepine) antihyperlipidemic drugs. Examples of benzothiazepine antihyperlipidemics for use in the combination therapy of the present invention can be found in U.S. patent No. 5,994,391, which is incorporated herein by reference. Certain benzothiazepine * antihyperlipidemic agents are described in WO 93/16055. In addition, the antihyperlipidemic or hypocholesterolemic agents used in combination with the compounds of the present invention may be HMG Co-A reductase inhibitors. Examples of HMG Co-A reductase inhibitors for use in the combination therapy of the present invention include, respectively, benfluvastatin, fluvastatin, lovastatin, provastatin, simvastatin, atorvastatin, cerivastatin, bervastatin, ZD-9720 (described in PCT patent application No. WO97/06802), ZD-4522(CAS No. 147098-20-2 for the calcium salt; CAS No. 147098-18-8 for the sodium salt; described in European patent No. EP 521471), BMS 180431(CAS No. 129829-03-4), or NK-104(CAS No. 141750-63-2). Combination therapy of the compounds of the present invention with antihyperlipidemic or hypocholesterolemic agents may be useful, for example, in reducing the risk of atherosclerotic lesions forming in blood vessels. For example, atherosclerotic lesions often initially occur at sites of inflammation in blood vessels. Antihyperlipidemic or hypocholesterolemic drugs have been identified to reduce the risk of atherosclerotic lesions forming by lowering lipid levels in the blood. The present invention is not limited to a single mechanism of action, and it is believed that the combination of the compounds of the present invention will collectively provide more effective control of atherosclerotic lesions, for example by lowering blood lipid levels, thereby reducing inflammation in the blood vessels.

In another embodiment of the invention, the compounds of the invention are useful in the treatment of central nervous disorders or diseases such as migraine in combination with other compounds or drugs. For example, the compounds may be used in combination with caffeine, a 5-HT-1B/1D agonist (e.g., triptan such as sumatriptan, naratriptan, zolmitriptan, rizatriptan, almotriptan or frovatriptan), a dopamine D4 antagonist (e.g., sonepprazole), aspirin, acetaminophen, ibuprofen, indomethacin, naproxen sodium, a combination therapy of methacin amine, chloraldoprine, butabiturate, ergot alkaloids (e.g., ergotamine, dihydroergotamine, bromocriptine, ergotamine, or methylergotamine), tricyclic antidepressants (e.g., amitriptyline or nortriptyline), 5-hydroxytryptamine antagonists (e.g., dimethylergotamine or cyproheptadine), beta-andrenegical antagonists (e.g., propranolol, timolol, atenolol, nadolol, or metoprolol), or monoamine oxidase inhibitors (e.g., phenelzine or isocarboxazid).

Another embodiment provides a combination therapy of a compound of the present invention together with an opioid compound. Can be used for the combination therapyOpioid compounds of (a) include, but are not limited to, morphine, methadone, hydromorphone, oxymorphone, levorphanol, allyllevorphanol, codeine, dihydrocodeine, dihydrohydroxycodeine, pentazocine, hydrocodone, oxycodone, nalmefene, etorphine, levorphanol, fentanyl, sufentanil, DAMGO, butorphan, buprenorphine, naloxone, naltrexone, CTOP, diprenorphine, beta-funaltrexanine, naloxonazine, allylmorphine, pentazocine, nalbuphine, naloxonone, benzoylhydrazone, bremazocine, ethylketocycloazocin, U50,488, U69,593, spirolin, nor-binaltritorphine, naltrinine, PE, [ D-la²，glu⁴]deltorphin, DSLET, methionine-enkephalin, leucine-enkephalin, beta-endorphin, dynorphin A, dynorphin B and alpha-neoendorphin. An advantage of the combination of the present invention with an opioid compound is that the compounds of the present invention may reduce the dosage of the opioid compound, thereby reducing the risk of opioid side effects such as opioid addiction.

The term "alkyl", alone or in combination, refers to a straight or branched chain acyclic alkyl group preferably containing from 1 to about 10 carbon atoms, more preferably from 1 to about 6 carbon atoms. "alkyl" also includes cycloalkyl groups containing from 3 to about 7 carbon atoms, preferably from 3 to 5 carbon atoms. The alkyl group may be optionally substituted with groups as defined below. Examples of such groups include methyl, ethyl, chloroethyl, hydroxyethyl, n-propyl, isopropyl, n-butyl, cyanobutyl, isobutyl, sec-butyl, tert-butyl, pentyl, aminopentyl, isopentyl, hexyl, octyl and the like.

The term "alkenyl" refers to a straight or branched chain unsaturated acyclic hydrocarbon group containing at least one double bond. Such groups contain from 2 to about 6 carbon atoms, preferably from 2 to about 4 carbon atoms, more preferably from 2 to about 3 carbon atoms. Said alkenyl group may be optionally substituted with groups as defined below. Examples of suitable alkenyl groups include propenyl, 2-chloropropenyl, buten-1-yl, isobutenyl, penten-1-yl, 2-methylbuten-1-yl, 3-methylbuten-1-yl, hexen-1-yl, 3-hydroxyhexen-1-yl, hepten-1-yl, octen-1-yl and the like.

The term "alkynyl" refers to a straight or branched chain unsaturated acyclic hydrocarbon radical containing one or more triple bonds and from 2 to about 6 carbon atoms, preferably from 2 to about 4 carbon atoms, and more preferably from 2 to about 3 carbon atoms. Said alkynyl group may be optionally substituted with groups as defined below. Examples of suitable alkynyl groups include ethynyl, propynyl, hydroxypropynyl, butyn-1-yl, butyn-2-yl, pentyn-1-yl, pentyn-2-yl, 4-methoxypentyn-2-yl, 3-methylbutyn-1-yl, hexyn-2-yl, hexyn-3-yl, 3-dimethylbutyn-1-yl, and the like.

The term "alkoxy" includes straight or branched chain, oxygen-containing groups each having from 1 to about 6 carbon atoms, preferably from 1 to 3 carbon atoms, such as methoxy. The term "alkoxyalkyl" also includes alkyl groups that contain one or more alkoxy groups attached to the alkyl group, i.e., to form monoalkoxyalkyl and dialkoxyalkyl groups. Examples of such groups include methoxy, ethoxy, propoxy, butoxy and tert-butoxyalkyl. "alkoxy" may be further substituted with one or more halogen atoms, such as fluorine, chlorine or bromine, to provide "haloalkoxy". Examples of such groups include fluoromethoxy, chloromethoxy, trifluoromethyloxy, difluoromethyloxy, trifluoroethoxy, fluoroethoxy, tetrafluoroethoxy, pentafluoroethoxy and fluoropropoxy.

The term "alkylthio" includes groups containing a straight or branched chain alkyl group of 1 to about 6 carbon atoms attached to a divalent sulfur atom. An example of "lower alkylthio" is methylthio (CH)₃-S-)。

The term "alkylthioalkyl" includes an alkylthio group attached to an alkyl group. Examples of such groups include methylthiomethyl.

The term "halo" refers to a halogen such as a fluorine, chlorine, bromine or iodine atom.

The term "heterocyclyl" refers to a saturated or unsaturated mono-or polycyclic carbocyclic ring in which one or more carbon atoms are replaced by N, S, P or O. For example, it includes the following structures:

z, Z therein¹、Z²Or Z³C, S, P, O or N, provided that Z, Z¹、Z²Or Z³One is not carbon, but when attached to another Z atom by a double bond, or when attached to another O or S atom, it is not O or S. In addition, only when Z, Z¹、Z²Or Z³Optional substituents, each being C, are understood to be attached to Z, Z¹、Z²Or Z³The above. The term "heterocyclyl" also includes fully saturated ring structures such as piperazinyl, dioxanyl, tetrahydrofuranyl, oxiranyl, aziridinyl, morpholinyl, pyrrolidinyl, piperidinyl, thiazolidinyl, and others. The term "heterocyclyl" also includes partially unsaturated ring structures such as dihydrofuranyl, pyrazolinyl, imidazolinyl, pyrrolinyl, chromanyl, dihydrophenylsulfanyl, and others.

The term "heteroaryl" refers to a fully unsaturated heterocyclyl group.

In "heterocyclyl" or "heteroaryl", the point of attachment to the molecule may be on a heteroatom on the ring or elsewhere.

The term "cycloalkyl" refers to a mono-or polycyclic carbocyclic ring wherein each ring contains from 3 to about 7 carbon atoms, preferably from 3 to about 5 carbon atoms. Examples include cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloalkenyl, and cycloheptyl. The term "cycloalkyl" additionally includes spiro systems in which the cycloalkyl ring has a carbon ring atom in common with the 7-membered heterocyclic ring of benzothiazepine.

The term "oxo" refers to an oxygen bearing a double bond.

The term "alkoxy" refers to a group that includes an alkyl group attached to an oxygen atom, such as a methoxy group. More preferred alkoxy groups are "lower alkoxy" groups having from 1 to about 10 carbon atoms. More preferred alkoxy groups have 1 to about 6 carbon atoms. Examples of such groups include methoxy, ethoxy, propoxy, isopropoxy, butoxy and tert-butoxy.

The term "aryl" refers to a fully unsaturated mono-or polycyclic carbocyclic ring including, but not limited to, substituted or unsubstituted phenyl, naphthyl, or anthracenyl.

The term "combination therapy" refers to the treatment of a disorder or disease described in the present disclosure with two or more therapeutic agents, e.g., atherosclerosis, pain, inflammation, migraine, neoplasia, angiogenesis-related conditions or diseases, and the like. Such administration includes substantially simultaneous co-administration of these therapeutic agents, for example, in a single capsule with a fixed ratio of active ingredients or in multiple separate capsules each containing an active ingredient. In addition, such administration also includes the use of various types of therapeutic agents in a sequential manner. In either case, the treatment provides beneficial pharmaceutical combinations for treating the conditions or diseases described herein.

The phrase "therapeutically effective" means that the combined amounts of the active ingredients in the combination therapy are such that the desired combined amount is achieved. The combined amounts will achieve the goal of reducing or eliminating hyperlipidemia.

In one embodiment, the invention provides a compound, or salt thereof, having a structure corresponding to formula (la)1The structure of (1):

in-situ type1In which X is selected from the group consisting of-S-, -S (O) -and-S (O)₂-. X is preferably-S-. R²Is selected from C₁-C₆Alkyl radical, C₂-C₆Alkenyl radical, C₂-C₆Alkynyl, C₁-C₅alkoxy-C₁Alkyl and C₁-C₅alkylthio-C₁Alkyl, wherein each of these groups is optionally substituted with one or more substituents selected from-OH, alkoxy, and halogen.R²Preferably C substituted by a substituent selected from the group consisting of-OH, alkoxy and halogen₁-C₆An alkyl group. As for R³And R⁸，R⁸Is selected from-OR¹⁴and-N (R)¹⁵)(R¹⁶) And R is³Selected from the group consisting of-H, -OH, -C (O) -R¹⁷、-C(O)-O-R¹⁸and-C (O) -S-R¹⁹(ii) a Or R⁸is-N (R)²⁰) -and R³is-C (O) -, wherein R³And R⁸Form a ring together with the atoms to which they are attached; or R⁸is-O-and R³is-C (R)²¹)(R²²) -, wherein R³And R⁸And together with the atoms to which they are attached form a ring. If R is³is-C (R)²¹)(R²²) -, then R⁴is-C (O) -O-R²³(ii) a Otherwise, R⁴is-H. R¹、R⁵、R⁶And R⁷Independently selected from-H, halogen, C₁-C₆Alkyl radical, C₂-C₆Alkenyl radical, C₂-C₆Alkynyl and C₁-C₅alkoxy-C₁An alkyl group. R⁹And R¹⁰Independently selected from-H, C₁-C₆Alkyl radical, C₂-C₆Alkenyl radical, C₂-C₆Alkynyl and C₁-C₅alkoxy-C₁An alkyl group; as for R¹¹And R¹²，R¹¹Selected from the group consisting of-H, -OH, -C (O) -O-R²⁴and-C (O) -S-R²⁵And R is¹²Selected from the group consisting of-H, -OH, -C (O) -O-R²⁶and-C (O) -S-R²⁷(ii) a Or R¹¹is-O-and R¹²is-C (O) -, wherein R¹¹And R¹²Form a ring together with the atoms to which they are attached; or R¹¹is-C (O) -, and R¹²is-O-, wherein R is¹¹And R¹²Form a ring together with the atoms to which they are attached. R¹³Is C₁An alkyl group. R₁₄Is selected from-H and C₁-C₆Alkyl, wherein when R¹⁴Is C₁-C₆When alkyl, R¹⁴Optionally substituted with one or more groups selected from cycloalkyl, heterocyclyl, aryl and heteroaryl. As for R¹⁵And R¹⁶，R¹⁵Selected from the group consisting of-H, alkyl and alkoxy, and R¹⁶Selected from the group consisting of-H, -OH, alkyl, alkoxy, -C (O) -R^27a、-C(O)-O-R²⁸and-C (O) -S-R²⁹(ii) a Wherein when R is¹⁵And R¹⁶When independently is alkyl or alkoxy, R¹⁵And R¹⁶Independently optionally substituted with one or more groups selected from cycloalkyl, heterocyclyl, aryl and heteroaryl; or R¹⁵is-H; and R is¹⁶Selected from the group consisting of cycloalkyl, heterocyclyl, aryl and heteroaryl. R¹⁷、R¹⁸、R¹⁹、R²⁰、R²¹、R²²、R²³、R²⁴、R²⁵、R²⁶、R²⁷、R^27a、R²⁸And R²⁹Independently selected from-H and alkyl, optionally substituted with one or more groups selected from cycloalkyl, heterocyclyl, aryl and heteroaryl; when R is¹、R²、R³、R⁴、R⁵、R⁶、R⁷、R⁸、R⁹、R¹⁰、R¹¹、R¹²、R¹³、R¹⁴、R¹⁵、R¹⁶、R¹⁷、R¹⁸、R¹⁹、R²⁰、R²¹、R²²、R²³、R²⁴、R²⁵、R²⁶、R²⁷、R^27a、R²⁸And R²⁹When any of (a) is independently a group selected from alkyl, alkenyl, alkynyl, alkoxy, alkylthio, cycloalkyl, heterocyclyl, aryl and heteroaryl, then the group is optionally substituted with one or more substituents selected from-OH, alkoxy and halogen.

In a preferred embodiment, R⁸is-OH. When R is⁸is-OH, X is preferably S. In another embodiment, R¹、R⁵、R⁶、R⁷、R⁹And R¹⁰Is independently selected from-H and C₁-C₃An alkyl group. R⁵、R⁶、R⁷、R⁹、R¹⁰Each is preferably H. R¹³Can be various groups, such as fluoromethylOr a methyl group. R¹May be C optionally substituted by a substituent selected from-OH and halogen₁-C₆An alkyl group; r¹Preferably C optionally substituted by halogen₁An alkyl group; more preferably, R¹Selected from the group consisting of fluoromethyl, hydroxymethyl and methyl. In an important embodiment, R¹May be a methyl group. In addition, R¹May be a fluoromethyl group. R¹Is hydroxymethyl. In another embodiment, R²Is C optionally substituted by one substituent selected from-OH, alkoxy and halogen₁-C₆An alkyl group. In a preferred embodiment, R²Is C optionally substituted by halogen₁An alkyl group. For example, R²May be a methyl group. In addition, R²May be a fluoromethyl group. In yet another embodiment, R²May be a hydroxymethyl group. In another embodiment, R²May be methoxymethyl.

In the compounds of the invention, R³、R⁴、R¹¹And R¹²Each is preferably-H. In this embodiment, R is further preferred¹、R⁵、R⁶、R⁷、R⁹And R¹⁰Independently selected from-H and C₁-C₃An alkyl group. R⁵、R⁶、R⁷、R⁹、R¹⁰Each is preferably-H. In this further embodiment, for example, R¹³May be fluoromethyl or in another embodiment R¹³May be a methyl group. In preferred compounds of these embodiments, R²Is C optionally substituted by a substituent selected from-OH, alkoxy and halogen₁-C₆An alkyl group. R²Preferably C optionally substituted by halogen₁An alkyl group. In one such embodiment, R²Is a fluoromethyl group. In another embodiment, R²Is methyl. Or, R²May be a hydroxymethyl group. Another option is R²May be methoxymethyl.

When R is¹³When it is methyl, R¹May be, for example, -H or optionally substituted by a substituent selected from-OH and halogenC₁-C₆An alkyl group. In a preferred embodiment R¹is-H. Or, R¹May be C optionally substituted by a substituent selected from-OH and halogen₁-C₆An alkyl group. For example, R¹Can be methyl, ethyl, n-propyl, isopropyl, n-butyl, sec-butyl, isobutyl, tert-butyl, pentyl isomers or hexyl isomers. For example, R¹May be an ethyl group. Or, R¹May be C optionally substituted by a substituent selected from-OH and halogen₁An alkyl group; for example, R¹May be a methyl group. Or, R¹May be a fluoromethyl group. In another alternative, R¹May be a hydroxymethyl group.

In another embodiment, R⁸Can be-OR¹⁴。R¹⁴As defined above. R¹⁴Preferably C optionally substituted by one or more groups selected from cycloalkyl, heterocyclyl, aryl and heteroaryl₁-C₆An alkyl group; r¹⁴More preferably C₁-C₃An alkyl group; r¹⁴Even more preferably methyl. In the compound1In another embodiment of (1), R⁸Can be-N (R)¹⁵)(R¹⁶) Wherein R is¹⁵And R¹⁶Is as defined above. In yet another embodiment, R⁸Can be-N (R)²⁰) -, and R³Can be-C (O) -, wherein R⁸And R³Form a ring together with the atoms to which they are attached. In another embodiment, R⁸Can be-O-and R³Can be-C (R)²¹)(R²²) -, wherein R⁸And R³Form a ring together with the atoms to which they are attached.

In-situ type1In the compound, R¹¹Can be selected from-OH, -C (O) -O-R²⁴and-C (O) -S-R²⁵。R¹¹preferably-OH. In another embodiment, when R¹¹When it is-OH, R¹²is-H.

However, the present invention also provides useful formulae1A compound of formula (I) wherein R¹¹is-O-and R¹²is-C (O) -, wherein R¹¹And R¹²Form a ring together with the atoms to which they are attached. In another useful embodiment, R¹¹is-C (O) -, and R¹²is-O-wherein R¹¹And R¹²Form a ring together with the atoms to which they are attached. Or, R¹²Can be selected from-OH, -C (O) -O-R²⁶and-C (O) -S-R²⁷. In this option, R¹¹preferably-H.

The invention also provides1A pharmaceutically acceptable salt of the compound. For example, such pharmaceutically acceptable salts can be those wherein the compound of the invention is in the form of a cation with at least one anionic counterion. Examples of anionic counterions for the pharmaceutically acceptable salts of the present invention include halides, carboxylates, sulfonates, sulfates, phosphates, phosphonates, resin-bound anions, oxides or nitrates. When the anionic counterion is a halide, for example, it may be fluoride, chloride, bromide or iodide. The halide counterion is preferably chloride. When the anionic counterion is a carboxylate (i.e., the anionic form of the carboxylic acid functional group-containing compound), the carboxylate counterion can vary widely. For example, the carboxylate counterion may be formate, acetate, propionate, trifluoroacetate, succinate, salicylate, DL-aspartate, D-aspartate, L-aspartate, DL-glutamate, D-glutamate, L-glutamate, glycerate, succinate, stearate, DL-tartrate, D-tartrate, L-tartrate, (+ -) -mandelate, (R) - (-) -mandelate, (S) - (+) -mandelate, citrate, mucate, maleate, malonate, benzoate, DL-malate, D-malate, L-malate, hemi-malate, 1-rhabdoacetate, 1-rhabdocarboxylate, propionate, lactate, acetate, xanthate, sulfoacetate, (+ -) -lactate, L- (+) -lactate, D- (-) -lactate, pamoate, D-alpha-galacturonate, glycerate, DL-ascorbate, D-ascorbate, L-ascorbate, DL-cystine, D-cystine, L-cystine, DL-homocystine, D- (-) -lactate, L- (+) -lactate, D- (-) -lactate, pamoate, D- (-) -galacturonate, glycerate, DL-ascorbate, D-cystine, L-homocystine, D-homocystine,l-homocystine salt, DL-cysteine salt, D-cysteine salt, L-cysteine salt, (4S) -hydroxy-L-prolinate salt, cyclopropane-1, 1-carboxylate salt, 2-dimethylmalonate salt, squarate salt, tyrosine anion, proline anion, fumarate salt, 1-hydroxy-2-naphthoate salt, phosphonoacetate salt, carbonate salt, bicarbonate salt, 3-phosphonopropionate salt, DL-pyroglutamate salt, D-pyroglutamate salt or L-pyroglutamate salt. Alternatively, the anionic counterion can be a sulfonate. For example, the sulfonate counterion can be a mesylate, tosylate, besylate, triflate, esylate, (±) -camphorsulfonate, naphthalenesulfonate, 1R- (-) -camphorsulfonate, 1S- (+) -camphorsulfonate, 2-mesitylsulfonate, 1, 5-naphthalenedisulfonate, 1, 2-ethanedisulfonate, 1, 3-propanedisulfonate, 3- (N-morpholino) propanesulfonate, biphenylsulfonate, isethionate, or 1-hydroxy-2-naphthalenesulfonate. In another embodiment, the anionic counterion can be a sulfate. Examples of sulfates useful in the present invention include, but are not limited to, sulfate, potassium bisulfate, sodium bisulfate, and sulfuric acid. The anionic counterion may be a sulfamate. When the anionic counterion is a phosphate, it can be, for example, phosphate, monobasic phosphate, potassium dibasic phosphate, dipotassium phosphate, potassium phosphate, sodium hydrogen phosphate, disodium phosphate, sodium phosphate, monocalcium phosphate, calcium phosphate, dibasic calcium phosphate, tribasic calcium phosphate (Ca)₃(PO₄)₂) Or a hexafluorophosphate salt. The anionic counterion can be a phosphonate. For example, the phosphonate counterion can be a vinyl phosphonate, 2-carboxyethyl phosphonate, or phenyl phosphonate. Alternatively, the anionic counterion may be a nitrate. The salts may also be derived from addition reactions of the compounds with oxides such as zinc oxide.

If desired, the anionic counterions can be bound to the polymeric resin. In other words, the anionic counterion can be a resin-bound anion. For example, the resin-bound anion can be a polyacrylate resin, wherein the resin contains anionic glycolate groups. An example of the polyacrylate resin used for the salt of the present invention is Bio-Rex 70 (manufactured by Bio-Rad). In an alternative example, the resin-bound anion can be a sulfonated poly (styrene divinylbenzene) copolymer resin. Non-limiting examples of sulfonated poly (styrene divinylbenzene) copolymer resins useful as anionic counterions in the present invention include Amberlite EPR-69 (Rohm & Haas) or Dowex50WX4-400 (Dow). If desired, the polyacrylate resin or sulfonated poly (styrene divinylbenzene) resin may be crosslinked with a crosslinking agent such as divinylbenzene.

In another embodiment, formula (II) is1A pharmaceutically acceptable salt of a compound can be a salt wherein a compound of the invention is in an anionic form with at least one cationic counterion. For example, the cationic counterion can be an ammonium cation, an alkali metal cation, an alkaline earth metal cation, a transition metal cation, or a resin-bound cation. When the cationic counterion is an ammonium ion, it may be substituted or unsubstituted. For example, the ammonium cation may be an alkylammonium cation or a di-, tri-or tetraalkylammonium cation. Additionally, the ammonium cation may be an aryl ammonium cation or a di, tri or tetraaryl ammonium cation. The ammonium cation may contain both alkyl and aryl groups. The ammonium cation may be aromatic, for example, a pyridinium cation. Other functional groups are also present in the form of ammonium cations. For example, the ammonium cation may be an ammonium, methylammonium, dimethylammonium, trimethylammonium, tetramethylammonium, ethanolammonium, dicyclohexylammonium, guanidinium, or ethylenediammonium cation. Alternatively, the cationic counterion can be an alkali metal cation such as a lithium cation, sodium cation, potassium cation, or cesium cation. In another alternative, the cationic counterion can be an alkaline earth metal cation such as a beryllium cation, a magnesium cation, or a calcium cation. If desired, the cation may be a transition metal cation such as a zinc cation.

If desired, the cationic counterion can be bound to the polymeric resin. In other words, the anionic counterion can be a resin-bound cation. For example, the resin-bound cation can be a cationically functionalized poly (styrene divinylbenzene) resin. An example of a cationically functionalized poly (styrene divinylbenzene) resin useful in the present invention is Bio-Rex-5(Bio-Rad), an ammonium functionalized resin. In another alternative, the resin-bound cation may be a cationically functionalized acrylic resin such as an aminated polyacrylic resin. An example of an aminated polyacrylic resin for use as a cationic counterion in the present invention is AG-4-XR (Bio-Rad).

In yet another embodiment of the invention, compounds of formula (I) are prepared by1The compounds may be present in zwitterionic form. In other words, the compound may contain both cationic and anionic sites within its molecule. Such zwitterionic form may exist as a counterion without isolation or it may exist as both a cationic and anionic counterion.

Another embodiment provides a method of treating or preventing a disease associated with inflammation, wherein the method comprises treating a patient in need thereof with a compound or salt of the invention in an amount to treat or prevent the disease associated with inflammation.

In another embodiment, the invention provides a compound of formula (I)21Method of compound or salt thereof:

wherein: x is selected from the group consisting of-S-, -S (O) -and-S (O)₂-。R²Is selected from C₁-C₆Alkyl radical, C₂-C₆Alkenyl radical, C₂-C₆Alkynyl, C₁-C₅alkoxy-C₁Alkyl and C₁-C₅alkylthio-C₁An alkyl group; r³⁰Selected from the group consisting of-H, -OH, -C (O) -R¹⁷、-C(O)-O-R¹⁸and-C (O) -S-R¹⁹；R¹、R⁵、R⁶And R⁷Independently selected from-H, halogen, C₁-C₆Alkyl radical, C₂-C₆Alkenyl radical, C₂-C₆Alkynyl and C₁-C₅alkoxy-C₁An alkyl group; r⁹And R¹⁰Independently selected from-H, C₁-C₆Alkyl radical, C₂-C₆Alkenyl radical, C₂-C₆Alkynyl and C₁-C₅alkoxy-C₁An alkyl group; as for R¹¹And R¹²：

R¹¹Selected from the group consisting of-H, -OH, -C (O) -O-R²⁴and-C (O) -S-R²⁵And R is¹²Selected from the group consisting of-H, -OH, -C (O) -O-R²⁶and-C (O) -S-R²⁷(ii) a Or R¹¹is-O-and R¹²is-C (O) -, wherein R¹¹And R¹²Form a ring together with the atoms to which they are attached; or R¹¹is-C (O) -, and R¹²is-O-wherein R¹¹And R¹²Form a ring together with the atoms to which they are attached;

R¹³is C₁An alkyl group;

R¹⁷、R¹⁸、R¹⁹、R²⁴、R²⁵、R²⁶、R²⁷and R^27aIndependently selected from-H and alkyl optionally substituted with one or more groups selected from cycloalkyl, heterocyclyl, aryl and heteroaryl; when R is¹、R²、R⁴、R⁵、R⁶、R⁷、R⁹、R¹⁰、R¹¹、R¹²、R¹³、R¹⁴、R¹⁵、R¹⁶、R¹⁷、R¹⁸、R¹⁹、R²⁴、R²⁵、R²⁶、R²⁷And R^27aAny of which is independently a group selected from alkyl, alkenyl, alkynyl, alkoxy, alkylthio, cycloalkyl, heterocyclyl, aryl and heteroaryl, which group is then optionally substituted with one or more substituents selected from-OH, alkoxy and halogen. Preparation of the Compounds21Comprises using a compound having a formula corresponding to23An alkyl acetimidate of structure (or salt thereof):

the process has a formula22A diamine compound of structure (la) (or a salt thereof):

in-situ type23In, R³¹Is C₁-C₆An alkyl group. R¹¹Preferably selected from-H and-OH. For example, R¹¹May be-H. Or, R¹¹May be-OH. When R is¹¹When is-H or-OH, R¹³Preferably methyl or halomethyl. More preferably R¹³Is methyl. Also, when R is¹¹When is-H or-OH, R³¹Preferably C₁-C₃Alkyl, more preferably ethyl. In a preferred embodiment, the treatment of the diamine compound with the alkyl acetimidate may be carried out in the presence of a base. For example, the base can be hydrazine, a metal sulfide, a metal hydroxide, a metal alkoxide, an amine, a hydroxylamine, a metal hydride, a metal amide complex, or a basic resin. When the base is a basic resin, it can be, for example, a polymer-bound diazabicyclo [4.4.0]Dec-2-ene resin. For example, the basic resin may have a structure bearing a diazabicyclo [4.4.0] attached to a copolymer]Poly (styrene divinylbenzene) copolymer backbones of dec-2-ene. When the base is an amine, it is essentially any substituted or unsubstituted amine. For example, the amine may be 1, 5-diazabicyclo [4.3.0 ]]Non-5-ene; 1, 4-diazabicyclo [2.2.2]Octane; or 1, 8-diazabicyclo [5.4.0 ]]Undec-7-ene. When the base is an alkali metal hydroxide, it may be, for example, potassium hydroxide or sodium hydroxide. When the base is a metal hydride, it may be, for example, sodium hydride, potassium hydride or calcium hydride.

In another embodiment of the present invention, there is provided a process for preparing a compound having a structure corresponding to formula (la)22A process for the preparation of a diamine compound of the structure (or a salt thereof), wherein the process comprises treating a diamine compound having a structure corresponding to formula (la) 24A protected diamine compound of structure (or a salt thereof):

wherein R is³³Is selected from-NH₂And a protected amino group; r³²Is a protected amino group; and R¹⁴Is selected from-H and C₁-C₆An alkyl group; wherein when R is¹⁴Is C₁-C₆When alkyl, R¹⁴Optionally substituted with one or more groups selected from cycloalkyl, heterocyclyl, aryl and heteroaryl; wherein the treatment is carried out with a deprotecting agent, thereby producing the diamine compound. The protected amino groups used in the present invention vary widely in nature. Theodora W.Greene and Peter G.M.Wuts (organic synthetic protecting groups, 3 rd edition, John Wiley&Sons, New York, 1999, page 494-653) describes the pair R used in the present invention³²Or R³³A plurality of protected amino groups. For example, R³²Or R³³Each of which is or are both 4-chlorobenzylimino. When R is³³In the case of 4-chlorobenzylimino, R is preferably³²is-NH₂. In another embodiment, R³²Or R³³Can each or both be tert-butoxycarbonylamino. When R is³³In the case of tert-butoxycarbonylamino, R³²Is preferably-NH₂. In another embodiment, R³²Or R³³Each of which may be or may be N-phthalimido. When R is³³In the case of N-phthalimido, R is preferred³²is-NH₂. In another embodiment, R³²Or R³³Each of which may be or may be benzyloxycarbonylamino. In a preferred embodiment of the invention, the protected amino group is any such group resulting from the reaction of an aldehyde with the relevant amino group to form a schiff base. A wide variety of deprotecting agents can be advantageously used in the invention to facilitate24Is converted into22. Greene and Wuts describe a number of such deprotecting agents (see above). For example, when the protected amino group is 4-chlorobenzylimino or tert-butoxyIn the case of aminocarbonylamino, the deprotecting agent is preferably an acid. Some useful acid deprotecting agents include, but are not limited to, hydrochloric acid, hydrobromic acid, sulfuric acid, trifluoroacetic acid, phosphoric acid (phosphoric acid), and acetic acid. In another embodiment, when R³²Or R³³In the case of N-phthalimido, the deprotecting agent may be either an acid or a base. When the deprotecting agent for N-phthalimido group is a base, the base can be, for example, hydrazine, metal sulfides, metal hydroxides, metal alkoxides, amines, hydroxylamines, and metal amide complexes. When the deprotecting agent for N-phthalimido group is an acid, the acid may be, for example, hydrochloric acid, hydrobromic acid, hydroiodic acid, sulfuric acid, methanesulfonic acid, trifluoromethanesulfonic acid, toluenesulfonic acid, benzenesulfonic acid, trifluoroacetic acid, phosphoric acid (phosphoric acid) or acetic acid. Compound 24 is preferably treated with a deprotecting agent in the presence of water.

In the reaction, R¹⁴preferably-H. R³³Is preferably-NH₂Or a salt thereof. R²Preferably methyl. In another preferred embodiment, R¹、R⁵、R⁶、R⁷、R⁹And R¹⁰Each is-H. A more preferred embodiment is where R is³³Is an embodiment of a tert-butoxycarbonylamino group. In a particularly preferred embodiment, the compounds24(or salts thereof) having a formula corresponding to25The structure of (1):

wherein R in brackets means that the carbon in the alpha position with respect to the carboxylic acid function is in the absolute configuration of R. In other words, compound 25 is the R-enantiomer or a salt thereof.

The invention also provides a process for preparing protected diamine compounds24(or a salt thereof), wherein the method comprises treating the microorganism with a compound having formula (la)27Protected aminoethyl alkylated compounds of structure

Wherein R is³⁴For nucleophilic substitution of the leaving group, treatment having formula26A sulfhydryl compound of the structure:

thereby forming a protected diamine compound. The reaction is preferably carried out in the presence of a base. For example, the base can be hydrazine, metal sulfides, metal hydroxides, metal alkoxides, amines, hydroxylamines, and metal amide complexes. The base is preferably an alkali metal hydroxide, more preferably potassium hydroxide or sodium hydroxide. Formula (II)27R in the structure³⁴Can vary over a wide range and can represent essentially any nucleophilic leaving group that generates or can be converted to a pharmaceutically acceptable anion. That is, (R)³⁴)^-An anion that is a pharmaceutically acceptable anion or an anion that can be converted to a pharmaceutically acceptable anion. For example, R³⁴Can be chloro, bromo, iodo, mesylate, tosylate, besylate, or triflate. R³⁴Preferably chloro, bromo or iodo, R³⁴More preferably bromine. In this reaction, R³³Is preferably-NH₂. In this reaction R²May be C optionally substituted by one or more substituents selected from-OH, alkoxy and halogen₁-C₆An alkyl group. R²Preferably C optionally substituted by one or more substituents selected from-OH, alkoxy and halogen₁-C₃Alkyl, more preferably, R²Is C₁-C₃An alkyl group. For example, R²Advantageously methyl may be used. In another embodiment, R⁵R of (A) to (B)⁶Each may be-H. In yet another embodiment, R¹And R⁷Can be independently selected fromH and C optionally substituted by one or more halogens₁-C₆An alkyl group; preferably R¹And R⁷Each is-H. In the present invention, R is preferred³²May be selected from 4-chlorobenzylimino, tert-butoxycarbonylamino and N-phthalimido, more preferably R³²Is tert-butoxycarbonylamino.

The invention also provides a preparation method of the compound28A method of making a sulfhydryl compound of structure:

wherein R is²、R⁵And R⁶As defined above, the process comprises treating a compound having the formula29A thiazolidine compound of structure:

wherein R is³⁵Is selected from-H and C₁-C₆A group of alkyl groups; thereby producing a hydrogen sulfide compound. The hydrolysis conditions preferably comprise contacting the thiazolidine compound with an acid in the presence of water. Preferably, the acid is selected from the group consisting of hydrochloric acid, hydrobromic acid, hydroiodic acid, sulfuric acid, methanesulfonic acid, trifluoromethanesulfonic acid, toluenesulfonic acid, benzenesulfonic acid, trifluoroacetic acid, phosphoric acid (phosphoric acid) and acetic acid. In one embodiment of the invention, R⁵And R⁶Each is-H. In another embodiment, the compounds29R of (A) to (B)²Is C optionally substituted by one or more substituents selected from-OH, alkoxy and halogen₁-C₆An alkyl group. R²Preferably C optionally substituted by one or more substituents selected from alkoxy and halogen₁-C₃Alkyl, more preferably R²Is C₁-C₃An alkyl group. For example, R²May be a methyl group. In another embodiment of the invention, the compounds29R of (A) to (B)³⁵Is methyl.

The invention also provides a preparation method of the compound30A methylthiazolidine compound of structure (or a pharmaceutically acceptable salt thereof)

Wherein R is³⁶Is C₁-C₆An alkyl group; the method comprises treating a deprotonatable compound having the formula31A thiazolidine compound of structure:

thereby producing the methylthiazolidine compound. Preferably, the methylation conditions comprise treating the deprotonatable thiazolidine compound with a base or methylating agent. The nature of the base can vary within wide limits. For example, the base can be a metal hydroxide, metal alkoxide, metal hydride, and metal amide complex. The base is preferably a metal amide complex. Partial metal amide complexes useful as bases in the present invention include, but are not limited to, lithium hexamethyldisilazide (hexamethyldisilazide), sodium hexamethyldisilazide, potassium hexamethyldisilazide, lithium diisopropylamide, sodium diisopropylamide, potassium diisopropylamide, sodium amide, lithium amide, potassium amide, sodium diethylamide, lithium diethylamide, potassium diethylamide, methyllithium, tert-butyllithium, sec-butyllithium, methylsodium, tert-butylsodium, sec-butylsodium, and methylmagnesium bromide. In the methylation reaction, R³⁶Can be C₁-C₃An alkyl group; for example R³⁶May be a methyl group.

The invention further provides a process for the preparation of deprotonatable thiazolidine compounds31Wherein the method comprises contacting cysteine C with pivalaldehyde under fusing conditions₁-C₆Alkyl ester to produce a deprotonatable thiazolidine compound. Preferred fusing conditions include contacting in the presence of a base. The base can vary within a wide range. For example, the base can be, but is not limited to, hydrazine, metal sulfides, metal hydroxides, metal alkyl bases, metal alkoxides, amines, hydroxylamines, and metal amide complexes. When the base is a metal amide complex, it may be, for example, lithium bis (trimethylsilyl) amide. In the fusion reaction, R³⁶Preferably C₁-C₃Alkyl radical, R³⁶More preferably methyl.

Another embodiment of the present invention provides a process for preparing a compound of formula32Method for producing alpha-amino acid compound of structure (or salt, enantiomer or racemate thereof)

Wherein: r²Is selected from C₁-C₆Alkyl radical, C₂-C₆Alkenyl radical, C₂-C₆Alkynyl, C₁-C₅alkoxy-C₁Alkyl and C₁-C₅alkylthio-C₁An alkyl group; r¹、R⁵、R⁶And R⁷Independently selected from-H, halogen, C₁-C₆Alkyl radical, C₂-C₆Alkenyl radical, C₂-C₆Alkynyl and C₁-C₅alkoxy-C₁An alkyl group; r⁹And R¹⁰Independently selected from-H, C₁-C₆Alkyl radical, C₂-C₆Alkenyl radical, C₂-C₆Alkynyl and C₁-C₅alkoxy-C₁An alkyl group; and when R is¹、R²、R⁵、R⁶、R⁷、R⁹And R¹⁰When any of them is independently a group selected from alkyl, alkenyl and alkynyl, said group is optionally substituted with one or more substituents selected from-OH, alkoxy and halogen; wherein the method comprisesUnder hydrolysis conditions treating a compound of formula33Hydantoin compounds of structure (or salts, enantiomers, or racemates thereof)

Thereby producing said alpha amino acid compound. For example, the hydrolysis conditions may comprise contacting hydantoin with an acid to produce an acid hydrolysate. Acids used for the hydrolysis reaction include, for example, hydrochloric acid, hydrobromic acid, hydroiodic acid, sulfuric acid, trifluoroacetic acid, or phosphoric acid. Alpha amino acid compound32The preparation method of (1) may further comprise treating the hydrolysate of the acid with an ion exchange resin. Alternatively, the hydrolysis conditions may comprise contacting the hydantoin compound with a base to produce a base hydrolysis product. Bases used for the alkaline hydrolysis include, but are not limited to, hydrazine, metal sulfides, metal hydroxides, or metal alkoxides. Compounds are preferred, whether base-catalyzed or acid-catalyzed hydrolysis33R in the structure¹、R⁵、R⁶And R⁷Each is-H. Also preferred is R⁹And R¹⁰Each is-H. In a particularly preferred embodiment, the α -amino acid compound (or a salt, enantiomer or racemate thereof) has the formula34The structure of (1):

the invention also provides a preparation method of the compound35A method of hydantoin compounds of structure (or salts, enantiomers, or racemates thereof):

wherein R is¹、R²、R⁵、R⁶、R⁷、R⁹And R¹⁰As defined above, wherein said process comprises contacting a cyanide source in the presence of an ammonium carbonate source and water with a cyanide source36An α -thioxone compound of the structure:

thereby producing a hydantoin compound. The cyanide source may be, for example, hydrogen cyanide or a metal cyanide salt. When the cyanide source is a metal cyanide salt, the salt is preferably sodium cyanide, potassium cyanide or lithium cyanide. More preferably, the metal cyanide salt is sodium cyanide. For compounds in the hydantoin-forming reaction36，R¹、R⁵、R⁶And R⁷Each is preferably-H. More preferably R⁹And R¹⁰Each is-H. Preparation of the Compounds35The method of (3) may further comprise a chiral separation step. When preparing the compound35When the process of (a) further comprises a chiral separation step, then the hydantoin compound product preferably has a compound33(or a salt or enantiomer thereof).

The present invention also provides a process for producing an alpha-thioxolone compound36Wherein the process comprises contacting a compound having the formula (la) with di-tert-butyl carbonate in the presence of a base37An aminothiol compound of the structure:

to produce an intermediate mixture; and reacting the intermediate mixture with a compound of formula38An α -chloroketone compound of the structure:

contact to generate alpha-a thioketone compound. The base in the reaction can vary widely. For example, the base can be, but is not limited to, hydrazine, metal sulfides, metal hydroxides, metal alkoxides, amines, hydroxylamines, and metal amide complexes. The base is preferably a metal hydroxide such as sodium hydroxide, potassium hydroxide or lithium hydroxide. In the reaction, R¹、R⁵、R⁶And R⁷Each is preferably-H. More preferably R⁹And R¹⁰Each is-H.

The term "pharmaceutically acceptable salts" includes salts commonly used to form alkali metal salts and to form addition salts of a free acid or a free base. The nature of the salt is not critical as long as it is pharmaceutically acceptable. Pharmaceutically acceptable salts are particularly useful as the products of the process of the present invention due to their greater water solubility relative to the corresponding parent or neutral compound. Such salts must have a pharmaceutically acceptable anion or cation. Suitable pharmaceutically acceptable acid addition salts of the compounds of the present invention may be prepared from inorganic or organic acids. Examples of such inorganic acids are hydrochloric acid, hydrobromic acid, hydroiodic acid, nitric acid, carbonic acid, sulfuric acid, and phosphoric acid. Suitable organic acids include those from the aliphatic, cycloaliphatic, aromatic, arylaliphatic, heterocyclic, carboxylic and sulfonic classes, examples of which are formic, acetic, propionic, succinic, glycolic, gluconic, lactic, malic, tartaric, citric, ascorbic, glucoronic, maleic, fumaric, pyruvic, aspartic, glutamic, benzoic, anthranilic, methanesulfonic, salicylic, p-hydroxybenzoic, phenylacetic, mandelic, pamoic (pamoic), methanesulfonic, ethanesulfonic, benzenesulfonic, sulfanilic, stearic, cyclohexylsulfamic, alginic (algenic), galacturonic. Suitable pharmaceutically acceptable base addition salts of the compounds of the present invention include metal salts made from aluminum, calcium, lithium, magnesium, potassium, sodium and zinc or organic salts made from N, N' -dibenzylethylenediamine, choline, chloroprocaine, diethanolamine, ethylenediamine, meglumine (N-methylglucamine) and procaine. Pharmaceutically acceptable acid addition salts of the compounds of the invention which are suitable include, where possible, salts derived from inorganic acids, for example hydrochloric, hydrobromic, boric, fluoroboric, phosphoric, metaphosphoric, nitric, carbonic (including carbonate and bicarbonate anions), sulfonic and sulfuric acids, and organic acids, for example acetic, benzenesulfonic, benzoic, citric, ethanesulfonic, fumaric, gluconic, glycolic, isothionic, lactic, lactobionic, maleic, malic, methanesulfonic, trifluoromethanesulfonic, succinic, toluenesulfonic, tartaric and trifluoroacetic acids. Suitable pharmaceutically acceptable base salts include ammonium salts, alkali metal salts such as sodium and potassium salts, and alkaline earth metal salts such as magnesium and calcium salts. All of these salts can be prepared in a conventional manner from the corresponding conjugate base or acid of the compounds of the invention by reacting the appropriate acid or base with the conjugate base or acid, respectively, of the compound. Another pharmaceutically acceptable salt is a resin-bound salt.

Although it is possible for the compounds of the invention to be administered as starting chemicals, it is preferred to have them present as pharmaceutical compositions. According to another aspect, the present invention provides a pharmaceutical composition comprising a compound of the invention, or a pharmaceutically acceptable salt or solvate thereof, together with one or more pharmaceutically acceptable carriers thereof and optionally one or more other therapeutic ingredients. The carrier must be acceptable in the sense of being compatible with the other ingredients of the formulation and not deleterious to the recipient thereof.

The formulations include those suitable for oral, parenteral (including subcutaneous, intradermal, intramuscular, intravenous and intraarticular), rectal and topical (including dermal, buccal, sublingual and intraocular) administration, although the most suitable route may depend on, for example, the condition and disease of the recipient. The formulations may be conveniently presented in unit dosage form and may be prepared by any of the methods well known in the art of pharmacy. All methods include the step of bringing into association the compound of the invention, or a pharmaceutically acceptable salt or solvate thereof, with the carrier which constitutes one or more accessory ingredients. In general, the formulations are prepared by uniformly and intimately bringing into association the active ingredient with liquid carriers or finely divided solid carriers or both, and then, if necessary, shaping the product into the desired formulation.

Formulations of the present invention suitable for oral administration may be presented as discrete units such as capsules, cachets or tablets each containing a predetermined amount of the active ingredient; as a powder or granules; as a solution or suspension in an aqueous liquid or a non-aqueous liquid; or as an oil-in-water liquid emulsion or a water-in-oil liquid emulsion. The active ingredient may also be presented as a bolus (bolus), electuary or paste.

Tablets may be prepared by compression or molding, optionally together with one or more accessory ingredients. Compressed tablets may be prepared by compressing in a suitable apparatus the active ingredient in a free-flowing form such as a powder or granules, optionally mixed with a binder, lubricant, inert diluent, lubricant, surfactant or dispersing agent. Molded tablets are prepared by molding or otherwise preparing a mixture of the powdered compound moistened with an inert liquid diluent in a suitable apparatus. The tablets may optionally be coated or scored and may be formulated so as to provide slow or controlled release of the active ingredient therein.

Parenteral formulations include aqueous and non-aqueous sterile injection solutions which may contain antioxidants, buffers, bacteriostats and solutes which render the formulation isotonic with the blood of the recipient to which it is to be administered; and aqueous and non-aqueous sterile suspensions which may include suspending agents and thickening agents. The formulations may be presented in unit-dose or multi-unit dose containers, for example, sealed ampoules and vials, and may be stored in a freeze-dried (lyophilized) condition requiring only the addition of the sterile liquid carrier, for example, saline, water for injections, immediately prior to use. Injections and suspensions may be prepared from sterile powders, granules and tablets of the kind previously described, ready for use.

Formulations for rectal administration may be presented as a suppository with a conventional carrier such as cocoa butter or polyethylene glycol.

Formulations for topical administration in the mouth, e.g., buccal or sublingual administration, include lozenges comprising the active ingredient in a flavoured base such as sucrose and acacia or tragacanth, and pastilles comprising the active ingredient in a base such as gelatin and glycerin or sucrose and acacia.

Preferred unit dosage formulations are those containing an effective amount of the active ingredient as described below, or an appropriate sub-dose thereof.

It will be understood that the formulations of the invention may include, in addition to the ingredients particularly mentioned above, other agents conventional in the art having regard to the type of formulation in question, for example, those suitable for oral administration may include flavoring agents.

The compounds of the invention may be administered orally or by injection at a dose of 0.001-2500mg/kg per day. The dosage range for adults is generally 0.005mg-10 g/day. Tablets or other forms provided in discrete units may conveniently contain an amount of a compound of the invention which is effective at such a dose or as multiple sub-doses of the same dose, e.g. units containing from 5mg to 500mg, usually from about 10mg to 200 mg.

The compounds of formula 1 are preferably administered orally or by injection (intravenously or subcutaneously). The exact amount of compound administered to a patient will be at the responsibility of the attending physician. However, the dosage employed will depend upon a number of factors including the age and sex of the patient, the precise disease to be treated and its severity. The route of administration also varies with the disease and its severity.

The compounds of the present invention may exist in tautomeric, geometric or stereoisomeric forms. The present invention contemplates all such compounds, including cis-and trans-geometric isomers, E-and Z-geometric isomers, R-and S-enantiomers, diastereomers, d-isomers, 1-isomers, racemic mixtures thereof, and other mixtures thereof, as falling within the scope of the present invention. Pharmaceutically acceptable salts of such tautomeric, geometric or stereoisomeric forms are also included within the scope of the invention.

The terms "cis" and "trans" refer to the form of geometric isomerism in which two carbon atoms connected by a double bond each have two higher (right linking) groups on the same side of the double bond (cis) or on opposite sides of the double bond (trans). Some of the compounds contain alkenyl groups and are intended to include cis and trans or "E" and "Z" geometric forms.

Some of the compounds contain one or more stereocenters and for each of the stereocenters R, S is intended to be included as well as mixtures of the R and S forms.

In carrying out the present invention, the following general synthetic procedures are useful.

Scheme 1a

Boc ═ tert-butoxycarbonyl

RT ═ room temperature

Et is ethyl

Me is methyl

Basic resin ═

Polymer-bound triazabicyclo [4.4.0] dec-5-ene

Scheme 1b

R is alkyl or alkoxyalkyl

(i) Pentane and dean-stark trap; (ii) HCO₂Na，Ac₂O，HCO₂H；

(iii)LiN[(CH₃)₃Si]₂，DMPU，THF，-78℃；

(iv) RI or R-SO₃CF₃(ii) a (v)6N HCl, reflux 2 days.

Scheme 1c

R is alkyl or alkoxyalkyl

(i)NaH，NMP，Boc-NHCH₂CH₂Br；(ii)1NHCl；

(iii) Ethyl acetimidate; (iv) ion exchange

Scheme 2

Scheme 3

R is alkyl

(i) Benzyl chloroformate, benzene; (ii) tosyl chloride, triethylamine; (iii) 2-methyl-L-cysteine-HCl, NaH, NMP; (iv) refluxing with 6N HCl; (v) ethyl acetimidate, NaOH; (vi) and (4) ion exchange.

Scheme 4

R is alkyl

(i)KHF₂，nBu₄NH₂F₃(ii) a (ii) Tosyl chloride, triethylamine;

(iii) 2-methyl-L-cysteine-HCl, NaH, NMP; (iv)6N HCl, reflux; (v) ethyl acetimidate, NaOH; (vi) and (4) ion exchange.

Scheme 5

R is alkyl or alkoxyalkyl

(i)CH₃OH, HCl; (ii) 4-chlorobenzaldehyde, (CH)₃CH₂)₃N，CH₃CN，MgSO₄；

(iii)NaN[(CH₃)₃Si]₂-78 deg.C; (iv) RI; (v) HCl; (vi) ethyl acetimidate, base; (vii) and (4) ion exchange.

Scheme 6

(i) HBr (Synthesis 1971646-7, J Chem Soc 19613448-52); (ii) k₂Cr₂O₇；(iii)Boc-NHCH₂CH₂SH, NaOH, toluene; (iv) NaCN, (NH)₄)₂CO₃EtOH; (v) chiral separation; (vi) 48% HBr; (vii) ethylidene amino acid ethyl ester base; (viii) HCl.

Scheme 7

Scheme 8

i) NaOH; ii) acetonitrile, reflux overnight; iii) diethyl carbonate, potassium tert-butoxide;

iv)1, 2-dibromoethane, DMF; v) NaOH, alpha-methyl cysteine ester

The following structures are illustrative of the many compounds provided by the present invention. The examples provided herein are not intended to be limiting and those skilled in the art will appreciate, in light of the present disclosure, that the present invention encompasses many alternative configurations.

The following compounds are examples of other compounds encompassed by the present invention or useful in the preparation of the compounds of the present invention.

(2R, 4R) -2-tert-butyl-1, 3-thiazoline-3-formyl-4-carboxylic acid methyl ester,

(2R, 4R) -2-tert-butyl-1, 3-thiazoline-3-formyl-4-methyl-4-carboxylic acid methyl ester,

S- [2- [ [ (1, 1-dimethylethoxy) carbonyl ] amino ] ethyl ] -2-methyl-L-cysteine trifluoroacetate, and,

(S) -1- [ (benzyloxycarbonyl) amino ] -2-propanol,

(S) -1- [ (benzyloxycarbonyl) amino ] -2-propanol tosylate,

S- [ (1R) -2- (benzyloxycarbonylamino) -1-methylethyl ] -2-methyl-L-cysteine trifluoroacetate salt,

S- [ (1R) -2-amino-1-methylethyl ] -2-methyl-L-cysteine hydrochloride,

S- (2-aminoethyl) -L-cysteine, methyl ester,

N- [ (4-chlorophenyl) methylene ] -S- [2- [ [ (4-chlorophenyl) methylene ] amino ] ethyl ] -L-cysteine, methyl ester,

N- [ (4-chlorophenyl) methylene ] -S- [2- [ [ (4-chlorophenyl) methylene ] amino ] ethyl ] -2-methyl-D/L-cysteine, methyl ester.

The following examples are provided to illustrate the present invention and are not intended to limit the scope of the present invention. Those skilled in the art will readily appreciate that variations in the conditions and procedures of the following preparative procedures can be used to prepare these compounds.

Example 1

S- [2- [ (1-iminoethyl) amino ] ethyl ] -2-methyl-L-cysteine, dihydrochloride

example-1A) (2R, 4R) -2-tert-butyl-1, 3-thiazoline-3-formyl-4-carboxylic acid methyl ester

See Jeanguenat and Seebach, J.chem.Soc.Perkin Trans.l, 2291(1991) and Pattenden et al Tetrahedron, 49, 2131 (1993): (R) -cysteine methyl ester hydrochloride (8.58g, 50mmol), neopentyl glycol were reacted with continuous removal of water using a dean Stark trap (Deau Stark trap)Aldehyde (8.61g, 100mmol) and triethylamine (5.57g, 55mmol) were refluxed in pentane (800 ml). The mixture was filtered and evaporated. The resultant thiazolidine (9.15g, 45mmol) and sodium formate (3.37g, 49.5mmol) were stirred in formic acid (68ml) and treated with acetic anhydride (13ml, 138mmol) dropwise over 1h at 0-5 ℃. The solution was warmed to room temperature and stirred overnight. The solvent was evaporated and washed with aqueous 5% NaHCO₃The neutralized residue was extracted with ether (3 times). The combined organic layers were dried (anhydrous MgSO)₄) Filtration and evaporation gave the title compound (8.65g) as white crystals crystallized from hexane-ether (80% overall yield, 8: 1 conformational isomer mixture).¹H NMR(CDCl₃) δ major conformer: 1.04(s, 9H), 3.29(d, 1H), 3.31(d, 1H), 3.78(s, 3H), 4.75(s, 1H), 4.90(t, 1H), 8.36(s, 1H). MSm/z (electrospray) 232(M + H)⁺(100％)，204(10)164(24)。

example-1B) (2R, 4R) -2-tert-butyl-1, 3-thiazoline-3-formyl-4-methyl-4-carboxylic acid methyl ester

At-78 ℃ and N₂Next, DMPU (25ml) was added to a solution of the product of example-1A (2R, 4R) -2-tert-butyl-1, 3-thiazoline-3-formyl-4-carboxylic acid methyl ester (8.65g, 37.4mmol) in anhydrous tetrahydrofuran (130ml), and the mixture was stirred for 5 minutes. 1M lithium bis (trimethylsilyl) amide (37.5ml) in tetrahydrofuran was added and the mixture was stirred for an additional 30 minutes. After addition of methyl iodide (5.84g, 41.1mmol), the mixture was maintained at-78 ℃ for 4 hours and then warmed to room temperature with constant stirring. The solvent was evaporated in vacuo and brine and ethyl acetate were added. The aqueous phase was extracted three times with EtOAc and then with 10% KHSO₄The combined organic layers were washed with water and brine. Then dried (anhydrous MgSO)₄) Filtered and stripped of all solvents under reduced pressure. Chromatography of the residual oil on silica, eluting with 1-10% EtOAc in hexanes, provided the title compound (5.78g, 63%, 2.4: 1 conformational isomer mixture).¹H NMR(CDCl₃) δ major conformer: 1.08(s, 9H), 1.77(s, 3H), 2.72(d, 1H), 3.31(d, 1H), 3.77(s, 3H), 4.63(s, 1H), 8.27(s, 1H). Minor conformers, 0.97(s, 9H), 1.79(s)3H), 2.84(d, 1H), 3.63(d, 1H), 3.81(s, 3H), 5.29(s, 1H), 8.40(s, 1H); MS M/z (electrospray) 246(M + H)⁺(100%), 188(55)160 (95). Retention time in daicel chemical industries chiralcel OAS column 16.5 minutes, 10-40% IPA hexane 0-25 minutes, > 95% ee.

example-1C) (2R) 2-methyl-L-cysteine hydrochloride

In N₂Next, the product of example-1B, (2R, 4R) -2-tert-butyl-1, 3-thiazoline-3-formyl-4-methyl-4-carboxylic acid methyl ester (5.7g, 23.2mmol) was stirred with 6N HCl (100ml) and vigorously refluxed for 2 days. The solution was cooled, washed with EtOAc and evaporated to give the product (2R) 2-methyl-cysteine hydrochloride (3.79g, 95%) as a pale yellow powder.¹H NMR(DMSO-d₆) δ 1.48(s, 3H)2.82(t, 1H), 2.96(bs, 2H), 8.48(s, 3H). MS M/z (electrospray) 136[ M + H⁺]。

example-1D) S- [2- [ [ (1, 1-Dimethylethoxy) carbonyl ] amino ] ethyl ] -2-methyl-L-cysteine trifluoroacetate

Sodium hydride (2.6g, 60% in mineral oil, 65mmol) was charged to an oven-dried, vacuum-cooled RB flask charged with oxygen-free 1-methyl-2-pyrrolidone (5 ml). The mixture was cooled to-10 ℃ and kept under N₂Stirring the mixture. The product, 2-methyl-L-cysteine hydrochloride from example-1C (3.6g, 21.0mmol) in oxygen-free 1-methyl-2-pyrrolidone (25ml) was added in portions. All of H₂After cessation of release, 2- [ (1, 1-dimethylethoxycarbonyl) -amino dissolved in oxygen-free 1-methyl-2-pyrrolidone (15ml) was added at-10 ℃ C]Ethyl bromide (4.94g, 21 mmol). The reaction was stirred for an additional 4 hours and warmed to room temperature. The solution was neutralized with 1N HCl and 1-methyl-2-pyrrolidone was removed by evaporation in vacuo. Reverse phase chromatography with 1-20% acetonitrile in 0.05% aqueous trifluoroacetic acid solution afforded the title compound (5.9g), which was recovered by lyophilization of the appropriate fractions.¹H NMR(DMSO-d₆/D₂O) δ 1.31(s, 9H), 1.39(s, 3H), 2.55(m, 2H), 2.78(d, 1H), 3.04(d, 1H), 3.06(t, 2H). To C₁₁H₂₂N₂O₄HRMS calculated value of S: 279.1375(M + H)⁺) Found 279.1379.

example-1E) S- (2-aminoethyl) -2-methyl-L-cysteine hydrochloride

The product of example-1D, S- [2- [ [ (1, 1-dimethylethoxy) carbonyl]Amino group]Ethyl radical]-2-methyl-L-cysteine trifluoroacetate (5.5g, 14.0mmol) was dissolved in 1N HCl (100ml) and stirred under nitrogen at room temperature overnight. Freeze-drying to remove the solution to obtain the target product S- (2-aminoethyl) -2-methyl-L-cysteine hydrochloride,¹H NMR(DMSO-d₆/D₂O)δ1.43(s，3H)，2.72(m，2H)，2.85(d，1H)，2.95(t，2H)，3.07(d，1H)。M/z[M+H⁺]179。

the product of example-1E was dissolved in H₂In O, the pH was adjusted to 10 with 1N NaOH, and ethyl acetimidate hydrochloride (1.73g, 14.0mmol) was added. The reaction was stirred for 15-30 minutes, the pH was raised to 10 and the process was repeated three times. The pH was adjusted to 3 with HCl and the solution was loaded onto a washed DOWEX50 WX4-200 column. By H₂O and 0.25MNH₄The column was washed with OH followed by 0.5M NH₄And (5) OH washing. Adding 0.5M NH₄The OH wash fractions were immediately frozen, combined and lyophilized to give an oil which was dissolved in 1N HCl and evaporated to give the title compound as a white solid (2.7 g).¹H NMR(DMSO-d₆/D₂O) δ 1.17(s, 3H), 2.08(s, 3H), 2.52(d, 1H), 2.68(m, 2H), 2.94(d, 1H), 3.23(t, 2H). To C₈H₁₈N₃O₂HRMS calculated value of S: 220.1120[ M + H⁺]Found 220.1133.

Example 2

2- [ [ [2- [ (1-iminoethyl) amino ] ethyl ] thio ] methyl ] -O-methyl-D-serine, dihydrochloride

The steps and methods used in this example are the same as those of example 1, except that methoxymethyl iodide is used instead of methyl iodide in the step of example-1B. These methods yielded the title product as a white solid (2.7 g).¹H NMR(D₂O) δ 2.06(s, 3H), 2.70(m, 3H), 3.05(d, 1H), 3.23(s, 3H), 3.32(t, 2H), 3.46(d, 1H), 3.62(d, 1H). To C₉H₂₀N₃O₃HRMS calculated value of S: 250.1225[ M + H⁺]Found 250.1228.

Example 3

S- [ (1R) -2- [ (1-iminoethyl) amino ] -1-methylethyl ] -2-methyl-L-cysteine, dihydrochloride

example-3A (S) -1- [ (benzyloxycarbonyl) amino ] -2-propanol

Benzyl chloroformate (10.23g, 60mmol) in dry benzene (120ml) was added slowly and portionwise over 20 minutes to a solution of (S) -1-amino-2-propanol (9.76g, 130mmol) in dry benzene (60ml) at 0 deg.C while stirring vigorously under nitrogen. The mixture was stirred at 0 ℃ for 1 hour, then allowed to warm to room temperature and stirred for an additional 2 hours. In the organic layer over anhydrous MgSO₄The mixture was washed with water (twice) and brine (twice) before drying. Evaporation of all solvents gave the title product as an oil.¹H NMR(CDCl₃) δ 1.22(d, 3H), 2.40(bs, 1H), 3.07(m, 1H), 3.37(m, 1H), 3.94(m, 1H), 5.16(s, 2H), 5.27(m, 1H), 7.38(m, 5H). MS rm/z (electrospray) 232[ M +23 ]]⁺(100％)，166(96)。

example-3B) (S) -1- [ (benzyloxycarbonyl) amino ] -2-propanol tosylate

To the product of example-3A, (S) -1 was added at 0 ℃ over a period of 20 minutes- [ (benzyloxycarbonyl) amino group]A solution of-2-propanol (9.74g, 46.7mmol) and triethylamine (7.27g, 72mmol) in dichloromethane (60ml) was added slowly and portionwise to toluoylxanthochloride (9.15g, 48mmol) in dichloromethane (18ml) with vigorous stirring under nitrogen. The mixture was warmed to room temperature and stirred under nitrogen for an additional 36 hours. Over anhydrous MgSO₄Before drying, 1N HCl, water, 5% NaHCO was used₃The organic layer was washed with solution, water and brine. Evaporation of all solvents gave a white solid which was passed through a silica column containing ethyl acetate/hexane (1: 4) to remove excess tosyl chloride, then eluted with ethyl acetate/hexane (1: 3) to give the title product as a white crystalline solid. These materials were recrystallized from ethyl acetate/hexane to give white needles (10.8 g).¹HNMR(CDCl₃) δ 1.22(d, 3H), 2.39(s, 3H), 3.20(m, 1H), 3.43(dd, 1H), 4.66(m, 1H), 5.02(m, 1H), 5.04(ABq, 2H), 7.34(m, 7H), 7.77(d, 2H). MS M/z (electrospray) 386[ M +23 [)]⁺(100%), 320 (66). The product was assayed in a Regis technologies Inc. Perkle solvent (R, R) β -GEM1 HPLC column using a mobile phase isopropanol/hexane and 10% isopropanol gradient for 5 minutes followed by 10-40% isopropanol for 25 minutes using UV and Laser Polarimeter detectors. Retention time main peak: 22.2 min, > 98% ee.

example-3C) s- [ (1R) -2- (benzyloxycarbonylamino) -1-methylethyl ] -2-methyl-L-cysteine trifluoroacetate

The product 2-methyl-L-cysteine hydrochloride from example-1C (1g, 6.5mmol) dissolved in oxygen-free 1-methyl-2-pyrrolidone (5ml) was added to oven-dried, N-washed₂In a purged round bottom flask, the system was cooled to 0 ℃. Sodium hydride (0.86g, 60% in mineral oil, 20.1mmol) was added and the mixture stirred at 0 ℃ for 15 min. The product of example-3B, (2S) -1- [ (N-benzyloxycarbonyl) amino) dissolved in oxygen-free 1-methyl-2-pyrrolidone (10ml) was added over 10 minutes]-2-propanol tosylate (2.5g, 7 mmol). After 15 minutes at 0 ℃, the reaction mixture was stirred at room temperature for 4.5 hours. The solution was acidified to pH 4 with 1N HCl and passed through vacuumThe 1-methyl-2-pyrrolidone was removed by evaporation. Reverse phase chromatography, eluting with 20-40% acetonitrile in 0.05% aqueous trifluoroacetic acid yielded the title compound (0.57g) recovered by lyophilization.¹H NMR(H₂O, 400MHz) δ 1.0(d, 3H), 1.4(s, 3H), 2.6(m, 2H), 2.8(m, 1H), 3.1(m, 2H), 3.6(s, 1H), 5.0(ABq, 2H), 7.3(m, 5H). MS M/z (electrospray) 327[ M + H⁺](100%), 238(20), 224(10) and 100 (25).

Example 3D) S- [ (1R) -2-amino-1-methylethyl ] -2-methyl-L-cysteine hydrochloride

The product of example-3C, S- [ (1R) -2- (benzyloxycarbonylamino) -1-methylethyl]-2-methyl-L-cysteine trifluoroacetate (0.5g, 1.14mmol) was dissolved in 6N HCl and refluxed for 1.5 h. The mixture was cooled to rt and extracted with EtOAc. The aqueous layer was concentrated in vacuo to give the title product (2R, 5R) -S- (1-amino-2-propyl) -2-methyl-cysteine hydrochloride (0.29g), which was used without further purification.¹H NMR(H₂O, 400MHz) δ 1.2(m, 3H), 1.4(m, 3H), 2.7(m, 1H), 2.8-3.2(m, 2H), 3.4(m, 1H), (somewhat bimodal due to rotameric forms). MS m/z (electrospray): 193[ M + H⁺](61%), 176(53), 142(34), 134(100) and 102 (10).

The product of example-3D, S- [ (1R) -2-amino-1-methylethyl]-2-methyl-L-cysteine hydrochloride (0.2g, 0.76mmol) was dissolved in 2ml H₂In O, the pH was adjusted to 10.0 with 1N NaOH, and Ethyleneimidate hydrochloride (0.38g, 3mmol) was added in four portions over 10 minutes, and the pH was adjusted to 10.0 with 1N NaOH if necessary. After 1 hour, the pH was adjusted to 3 with 1N HCl. The solution was loaded into a water washed DOWEX50 WX4-200 column. By H₂O and 0.5N NH₄The column was washed with OH. The basic fractions were combined and concentrated to dryness in vacuo. The residue was acidified with 1n hcl and concentrated to give the title compound of example 3 (49 mg).¹H NMR(H₂O, 400MHz) delta 1.3-1.0(m, 3H), 1.5(m, 3H), 2.1-1.8(m, 3H), 3.4-2.6(m, 5H), 3.6(m, 1H) (rotamers observed). MS m/z (electrospray): 234[ M + H ]⁺](100%), 176(10) and 134 (10))。

Example 4

S- [ (1S) -2- [ (1-iminoethyl) amino ] -1-methylethyl ] -2-methyl-L-cysteine, dihydrochloride

The procedure and method used herein were the same as in example 3, except that (R) -1-amino-2-propanol was used instead of (S) -1-amino-2-propanol in step example-3A, to give the objective substance S- [ (1S) -2- [ (1-iminoethyl) amino group]-1-methylethyl group]-2-methyl-L-cysteine hydrochloride.¹H NMR(H₂O, 400MHz) delta 3.6(m, 1H), 3.4-2.6(m, 5H), 2.1-1.8(m, 3H), 1.5(m, 3H) and 1.3-1.0(m, 3H). To C₉H₁₉N₃O₂S[M+H⁺]HRMS calculated of (a): 234.1276, found 234.1286.

Example 5

S- (1R/S) -2- [ (1-iminoethyl) amino ] -1-ethylethyl ] -2-methyl-L-cysteine, dihydrochloride

The procedure and method used for this synthesis are the same as for example 3, except that (R/S) -1-amino-2-butanol is used instead of (S) -1-amino-2-propanol in step example-3A.

Example 6

S- [ (1S) -2- [ (1-iminoethyl) amino ] -1- (fluoromethyl) ethyl ] -2-methyl-L-cysteine, dihydrochloride

In the presence of a catalyst₄NH₂F₃Treatment of 2- [ (2R) -Oxiranylmethyl with Potassium Hydrogendifluoride in the Presence of]A sample of (E) -1H-isoindole-1, 3-dione (G.Alexander et al Tetrahedron Asymmetry, 7, 1641-8, 1996) gave 2- [ (2R) -3-fluoro-2-hydroxypropyl]-1H-isoindole-1, 3-dione. The procedure and method used in this synthesis were the same as those of example 3, except that 2- [ (2R) -3-fluoro-2-hydroxypropyl was used]-1H-isoindole-1, 3-dione instead of (S) -1- [ (benzyloxycarbonyl) amino group in step example-3B]The title product was prepared in addition to 2-propanol.

Example 7

S- [2- [ (1-iminoethyl) amino ] ethyl ] -2-ethyl-L-cysteine, dihydrochloride

The procedure and method used for this synthesis were the same as those of example 1, except that ethyl trifluoromethanesulfonate was used instead of methyl iodide in example-1B. The title product was purified by reverse phase chromatography using a gradient of 10-40% acetonitrile in water (20% yield).¹H NMR(D₂O) δ 0.83(t, 3H), 1.80(m, 2H), 2.08(s, 3H), 2.68(m, 1H), 2.78(m, 1H), 2.83(m, 1H), 3.11(m, 1H), 3.36(t, 2H). To C₉H₂₀N₃O₂HRMS calculated value of S: 234.1276[ M + H⁺]Found 234.1284.

Example 8

S- [ (1R) -2- [ (1-iminoethyl) amino ] -1-methylethyl ] -2-ethyl-L-cysteine, dihydrochloride

The procedure and method used in this synthesis were the same as in example 1, except that ethyl trifluoromethanesulfonate was used instead of methyl iodide in example-1B. The 2-ethyl-L-cysteine hydrochloride thus obtained was treated as described in the procedures and methods of examples 3C-3E to give the title compound.

Example 9

S- [ (1R/S) -2- [ (1-iminoethyl) amino ] -1-ethylethyl ] -2-ethyl-L-cysteine, dihydrochloride

The procedure and method used for this synthesis were the same as those used in example 5, except that 2-ethyl-L-cysteine hydrochloride (prepared in example 7) was used instead of 2-methyl-L-cysteine hydrochloride, to give the title compound.

Example 10

S- [ (1S) -2- [ (1-iminoethyl) amino ] -1-fluoromethylethyl ] -2-ethyl-L-cysteine, dihydrochloride

The procedure and method used for this synthesis were the same as those used in example 6, except that (2R) -2-ethylcysteine hydrochloride (prepared in example 7) was used instead of (2R) -2-methylcysteine hydrochloride, to give the title compound.

Example 11

2- [ [ [ [2- (1-iminoethyl) amino ] ethyl ] thio ] methyl ] -D-valine, dihydrochloride example 11a) isopropyl trifluoromethanesulfonate

A stirred solution of silver triflate (25.25g, 98.3mmol) in diethyl ether (300ml) was treated with isopropyl iodide (16.54g, 98.5mmol) in diethyl ether (200ml) under nitrogen for 15 min. The mixture was stirred for 10 minutes and filtered. The filtrate was distilled under reduced pressure. The distillate was redistilled at atmospheric pressure to remove most of the ether, leaving a mixture of the title isopropyl trifluoromethanesulfonate-diethyl ether (84: 16 weight ratio) as a colorless liquid (15.64g, 70% after correction).¹H NMR(CDCl₃O, 400MHz) δ 1.52(d, 6H), 5.21 (heptad, 1H).

The procedure and procedure used herein were the same as those used in example 1, except that isopropyl trifluoromethanesulfonate was used instead of methyl iodide in example-1B. The crude title product was purified by reverse phase chromatography eluting with a gradient of 10-40% acetonitrile in water.¹H NMR(H₂O, 400MHz) δ 0.94(dd, 6H), 2.04 (heptad, 1H), 2.10(s, 3H), 2.65, 2.80(dm, 2H), 2.85, 3.10(dd, 2H), 3.37(t, 2H). To C₁₀H₂₂N₃O₂HRMS calculated value of S: 248.1433[ M + H⁺]Found 248.1450.

Example 12

2- [ [ [ (1R) -2- [ (1-iminoethyl) amino ] -1-methylethyl ] thio ] methyl ] -D-valine, dihydrochloride

The procedure and method used for this synthesis were the same as those used in example 3, except isopropyl trifluoromethanesulfonate (prepared in example-11A) was used instead of methyl iodide to give the title compound.

Example 13

2- [ [ [ (1R/S) -2- (1-iminoethyl) amino) -1-ethylethyl ] thio ] methyl ] -D-valine, dihydrochloride

The procedures and methods used for this synthesis were the same as those used in example 5, except that isopropyl (2R) -2-trifluoromethanesulfonate (prepared in example-11A) was used in place of methyl iodide to give the title compound.

Example 14

2- [ [ [ (1S) -2- [ (1-iminoethyl) amino ] -1-fluoromethylethyl ] thio ] methyl ] -D-valine, dihydrochloride

The procedure and method used for this synthesis were the same as those used in example 6, except that isopropyl (2R) -2-trifluoromethanesulfonate (prepared in example-11A) was used instead of methyl iodide to give the title compound.

Example 15

S- [ (R/S) -2- [ (1-iminoethyl) amino ] -1- (trifluoromethyl) ethyl ] -2-methyl-L-cysteine, dihydrochloride

Treatment of tert-butyl-N- (2-oxoethyl) carbamate with 1, 1, 1-trifluoroethylmagnesium bromide gave (R/S) -1- [ (1, 1-dimethylethoxycarbonyl) ] amino-4, 4, 4-trifluoro-2-butanol. The procedures and methods used for this synthesis were the same as those listed in example 3, except that (R/S) -1- [ (1, 1-dimethylethoxy) carbonyl ] amino-4, 4, 4-trifluoro-2-butanol was used instead of (S) -1- [ (benzyloxycarbonyl) amino ] -2-propanol, to give the title compound.

Example 16

S- [2- (1-iminoethylamino) ethyl ] -2-methyl- (D/L) -cysteine, bistrifluoroacetate salt

example-16A) S- (2-aminoethyl) -L-cysteine, methyl ester

A10 g (50mmol) sample of S- (2-aminoethyl) -L-cysteine was dissolved in 400mL of methanol. Anhydrous HCl was bubbled through the cooled solution for 30 minutes. After stirring overnight at room temperature, the solution was concentrated to give 12.7g of the title compound.

example-16B) N- [ (4-chlorophenyl) methylene ] -S- [2- [ [ (4-chlorophenyl) methylene ] amino ] ethyl ] -L-cysteine, methyl ester

A sample of the product S- (2-aminoethyl) -L-cysteine methyl ester from example-16A, 12.7g (50mmol) were dissolved in acetonitrile. To the solution was added 12.2g (100mmol) of anhydrous MgSO₄14g (100mmol) of 4-chlorobenzaldehyde and 100mmol of triethylamine. The mixture was stirred for 12 hours, concentrated to a small volume and diluted with 500mL of ethyl acetate. Continuous use of (0.1%) NaHCO₃The organic solution was washed with (2N) NaOH and brine solution. The organics were dried (anhydrous MgSO)₄) Filtration and concentration gave 7.5g of the title compound. [ M + H ]⁺]＝179。

example-16C) N- [ 4-chlorophenyl) methylene ] -S- [2- [ [ (4-chlorophenyl) methylene ] amino ] ethyl ] -2-methyl-D/L-cysteine methyl ester

The product N- [ (4-chlorophenyl) methylene of example-16B was treated with 17mmol of sodium bis (trimethylsilyl) amide in anhydrous THF at-78 deg.C under nitrogen]-S- [2- [ [ (4-chlorophenyl) group) Methylene group]Amino group]Ethyl radical]A sample of methyl-L-cysteine (7.5g, 17mmol) was treated with 2.4g (17mmol) of methyl iodide. The solution was kept at-78 ℃ for 4 hours and then heated to room temperature with continuous stirring. The solvent was evaporated in vacuo and brine and ethyl acetate were added. The aqueous phase was extracted three times with EtOAc and 10% KHSO₄The combined organic layers were washed with water and brine, and then dried (anhydrous MgSO)₄) Filtration and evaporation gave the title compound.

example-16D) S- (2-aminoethyl) -2-methyl-D/L-cysteine, hydrochloride

A sample (4.37g, 10mmol) of the product, N- [ 4-chlorophenyl) methylene ] -S- [2- [ [ (4-chlorophenyl) methylene ] amino ] ethyl ] -2-methyl-D/L-cysteine methyl ester from example-16C, was stirred and heated with 2N Cl overnight (60 deg.C) and the solution was washed three times with ethyl acetate. The aqueous solution was lyophilized to give the title compound.

A sample (2.5g, 10mmol) of the product S- (2-aminoethyl) -2-methyl-D/L cysteine dihydrochloride from example-16D was dissolved in H₂In O, the pH was adjusted to 10 with 1N NaOH. Ethylimidate hydrochloride (1.24g, 10.0mmol) was added to the reaction mixture. The reaction was stirred for 15-30 minutes and the pH was raised to 10 and the process was repeated three times. The pH was lowered to 4 with HCl solution and the solution was evaporated. The residue was purified by reverse phase HPLC using H containing 0.05% trifluoroacetic acid₂O as mobile phase gave the title product of example 16. M + H220.

Example 17

S- [2- [ (1-iminoethyl) amino ] ethyl ] -2-fluoromethyl-L-cysteine, dihydrochloride

Treatment of epibromohydrin with HF-pyridine to give dihalo-alcohol, with K₂Cr₂O₇Oxidizing the mixture to obtain the 1-bromo-3-fluoroacetone. In thatTreating the product with (1, 1-methylethoxy) -N- (2-thio (sulfonyl) ethyl) formamide in the presence of NaOH to obtain (1, 1-dimethylethoxy) -N- [2- (3-fluoro-2-oxopropylthio) ethyl]Formamide. Passing NaCN and (NH) through refluxing ethanol₄)₂CO₃Cyclizing it to racemic hydantoin and separating the enantiomers by chiral chromatography. The S-enantiomer was treated with hot 48% HBr solution to give S- (2-aminoethyl) -2-fluoromethyl-L-cysteine dihydrochloride, which was converted to the title compound by treatment with ethyl acetimidate in the presence of a base.

Example 18

(2R) -2-amino-3 [ [2- [ (1-iminoethyl) amino ] ethyl ] sulfinyl ] -2-methylpropanoic acid, dihydrochloride

The S- [2- [ (1-iminoethyl) amino group in 3ml of water]Ethyl radical]-2-methyl-L-cysteine, dihydrochloride salt (example 1, 0.2g, 0.73mmol) solution was stirred and cooled to 0 ℃ and 3% H was added in portions of 0.3ml₂O₂(0.8ml, 0.73mmol) of formic acid (0.4ml, 0.73 mmol). The cooling bath was removed and the reaction mixture was stirred at room temperature for 48 hours. The solution was concentrated in vacuo, diluted with water (10ml) and concentrated again to give the crude sulfone. The residue was chromatographed (C-18 reverse phase, using H containing 0.05% trifluoroacetic acid₂Mobile phase of O) to yield pure sulfone. The sulfone was treated with 1M HCl (10ml) and concentrated in vacuo to give 140mg of the two diastereomeric mixture of the title compound as a colorless oily HCl salt.¹H NMR(300MHz，D₂O) δ 1.5(s, 2H), 1.6(s, 1H), 2.0(s, 3H), 3.1(m, 2H), 3.3(m, 2H), 3.6(m, 2H). To C₈H₁₈N₃O₃HRMS calculated value of S: 236.1069[ M + H⁺]Found 236.1024.

Example 19

(2R) -2-amino-3 [ [2- [ (1-iminoethyl) amino ] ethyl ] sulfonyl ] -2-methylpropanoic acid dihydrochloride

The product S- [2- [ (1-iminoethyl) amino ] of example 1 is dissolved in 2ml of water]Ethyl radical]-2-methyl-L-cysteine, dihydrochloride salt (0.15g, 0.54mmol) solution was cooled to 0 deg.C and 3% H was added₂O₂(1.6ml, 1.46mmol) of formic acid (0.8ml, 14.6 mmol). The cooling bath was removed and the reaction mixture was stirred at room temperature for 18 hours. The solution was concentrated in vacuo, diluted with 10ml of water and concentrated again to give the crude sulfoxide. The residue was diluted with 4ml of water and the pH was adjusted to 9 with 2.5N NaOH. Acetone (5ml) was added followed by Boc₂O (0.2g), and the reaction was stirred at room temperature for 48 hours. The reaction mixture was adjusted to pH 6 with 1M HCl and concentrated in vacuo. Chromatography (C-18 reverse phase; 40-50% ACN: H)₂O, 0.05% TFA) residue to give pure Boc protected starting material. The fractions were concentrated in vacuo and the residue was treated with 1N HCl (3ml) for 1 h. The solution was concentrated to give 30mg of the title compound as a colorless oil.¹H NMR(400MHz，D₂O) Δ 4.0(d, 1H), 3.7(d, 1H), 3.6(t, 2H), 3.5(t, 2H), 2.1(s, 3H) and 1.5(s, 3H) ppm. To C₈H₁₈N₃O₄HRMS calculated value of S: 252.1018[ M + H⁺]Measured value 252.0992.

Example 20

S- [2- [ (1-iminoethyl) amino ] ethyl ] -2-methyl-L-cysteine

The DOWEX50 WX4-200(250g) in a glass chromatography column (38X 560mm) was washed with water until the eluate had a pH of 6. The product S-[2- [ (1-iminoethyl) amino group]Ethyl radical]-2-methyl-L-cysteine dihydrochloride (6g) solution was put into the column and washed with water to pH back to 6. Reuse 0.07M NH₄The column was washed with OH (flow rate about 15ml/min) and the alkaline fraction was immediately placed in a dry ice/acetone bath. The fractions were collected and concentrated to dryness by lyophilization to give the title compound.¹H NMR(400MHz，DMSOd₆) δ 3.4(m, 1H), 3.3(m, 1H), 3.0(d, 1H), 2.7(m, 1H), 2.4(m, 1H), 2.3(d, 1H), 2.1(s, 3H) and 1.1(s, 3H). To C₈H₁₇N₃O₂S+0.6H₂Analytical calculation of O: c41.76, H7.97, N18.26; found C41.43, H7.47, N17.96, Cl traces.

Example 21

S- [2- [ (1-iminoethyl) amino ] ethyl ] -2-methyl-L-cysteine, dihydrochloride

example-21A) (2R, 4R) -2-tert-butyl-1, 3-thiazoline-3-formyl-4-carboxylic acid methyl ester

The title material was prepared as described in j.chem.soc.perkin trans.1991, p 2291 and Tetrahedron 1993, p 2131. To a 2L RB flask equipped with a reflux condenser, dean-Stark trap, overhead stirrer, and thermocouple was added pivalaldehyde (23.7g, 0.275mole) dissolved in 700mL of toluene. Stirring was started and L-cysteine methyl ester hydrochloride (45g, 0.262mole) was added to the stirred solution. A stream of triethylamine (29.2g, 0.288moles) was added to the batch over a period of several minutes. The reaction mixture was heated to reflux and water was removed. The batch was heated for a total of 3 hours, cooled and filtered. The filter cake of the salt was washed with 250ml of fresh toluene and the washings combined. Formic acid (24.1g, 0.524moles) and solid sodium formate (19.6g, 0.288moles) were added and the resulting suspension was cooled to-5 ℃. Acetic anhydride (53.5g, 0.524moles) was carefully added to the mixture, maintaining the temperature of the batch below 5 ℃. After addition, the reaction was kept at room temperature with continued stirring for 18 hours, during which time the product precipitated out. The crude product was filtered and redissolved in 400ml EtOAc and filtered to remove the insoluble sodium salt. The organic solution was neutralized with 200ml of saturated sodium bicarbonate solution and the final aqueous layer pH was close to 7. The organic phase was separated and the aqueous layer was extracted with ethyl acetate. The organic layers were combined and concentrated to give the crude product (60.2g) as a viscous oil which slowly crystallized to a white solid. The solid was washed with 4% 2-propanol in cyclohexane to give 41.01g of the title product with a purity > 99.5% in a yield of 67.8% by GC. The desired cis isomer of the title product is present in an amount of more than 98%.

example-21B) (2R, 4R) -2-tert-butyl-1, 3-thiazoline-3-formyl-4-methyl-4-carboxylic acid methyl ester

Anhydrous lithium chloride (43.0g, 0.102moles) was mixed with 300ml of dimethoxyethane and 500ml of THF until a clear solution was obtained. A solution of the product of example-21A (50.0g, 0.216moles) in THF was added under a nitrogen atmosphere and cooled to-65 ℃. Methyl iodide (45.0g, 0.316moles) diluted with 45ml THF was added followed by 230ml of 1.0M THF lithium bis-trimethylsilyl amide solution. The reaction was stirred at-65 ℃ for 10 hours. The batch was quenched with 30g of acetic acid in 600ml of water. And extracted with 500ml of ethyl acetate. The organic layer was washed with saturated sodium bicarbonate and concentrated to give 51.22g (96%) of the title compound as a light brown solid.

example-21C) (2R) 2-methyl-L-cysteine hydrochloride

A sample (20g, 83mmoles) of the product of example-21B was placed in a round bottom flask equipped with an overhead stirrer and a reflux condenser. To this solid was added 100ml of concentrated hydrochloric acid. The reaction was heated slowly to 95 ℃ for 7 days. The reaction was treated with 250ml of toluene to remove non-polar organic impurities. The aqueous solution was then concentrated. The crude title product was obtained weighing 14g of orange resin. The resin was powdered in ether/dichloromethane and filtered to give 13g of the title material as a light brown hygroscopic powder.¹H NMR(D₂O) delta 4.70(s, HDO exchange), 3.08(d, 1H), 2.80(d, 1H), 1.48(s, 3H).

Example 21D)2- [ (1, 1-Dimethylethoxycarbonyl) -amino ] ethyl bromide

3 liters of ethyl acetate was placed in a 5L round bottom flask equipped with an overhead stirrer, thermocouple and nitrogen inlet and stirring was started. Di-tert-butyl dicarbonate (545g, 2.50moles) and 2-bromoethylamine hydrobromide (553.7g, 2.70moles) were added thereto under a nitrogen blanket. The reaction was cooled to 5 ℃ in an ice bath and N-methylmorpholine (273g, 2.70moles) was added dropwise over about half an hour. After the addition was complete, the batch was stirred overnight and warmed to room temperature. After 16 hours, it was quenched by the addition of 1.5L of deionized water. The organic layer was washed with dilute HCl, sodium bicarbonate solution, followed by brine. The solvent of the dried organic solution was removed to give an oil which was frozen to a pale yellow solid. The total amount of the title product obtained was 496g (88% yield), with a purity of about 96%.

example-21E) S- [2- [ [ (1, 1-Dimethylethoxy) carbonyl ] amino ] ethyl ] -2-methyl-L-cysteine acetate

435g of methanol was charged into an overhead stirrerThermocouple in 3L flask, and stored in N₂In the atmosphere. To the reaction flask, 150.0g (0.804moles) of the product of example-21C was carefully added while stirring to dissolve. A KOH solution prepared by dissolving 154.7g of solid KOH in 840ml of degassed methanol was added dropwise to the reaction solution, maintaining the temperature at 20-30 ℃. The product of example-21C (180.2g, 0.804moles) was dissolved in 375mL of methanol and the solution was added dropwise to the cold reaction mixture at 10-12 ℃ for an additional 1 hour. When the reaction is complete, the pH of the batch of reactants is adjusted to pH5. The reaction mixture was filtered through a pad of Celite and the filtrate was concentrated to give 454g of a brown solid product. The crude product was slurried with ethyl acetate to give 299g of an off-white solid. The crude title product solid was used without purification in the next step.¹H NMR as acetate (D)₂O)δ4.68(s，D₂O exchange), 3.12(m, 3H), 2.68(m, 3H), 1.83(s, 3H), 1.42(s, 3H), 1.32(s, 9H).

example-21F) S- (2-aminoethyl) -2-methyl-L-cysteine

To a round bottom flask equipped with an overhead stirrer and nitrogen sweep was added 150ml of 37% hydrochloric acid. The stirrer was started and 150ml of water were added to the vessel, followed by 173g of the crude product of example-21E. The reaction was stirred for two hours and the brown clear solution was concentrated to give the crude di-HCl salt of the title product (about 157g) as a brown syrup. It was redissolved in 200ml of water and decolorized with activated carbon. The solution was passed through a Dowex resin column and the neutral title product was eluted with aqueous ammonium hydroxide to give 64% pure material of about 94%.¹H MR(D₂O，300MHz)δ4.68(s，D₂O exchange), 2.9(m, 3H), 2.6(m, 3H), 1.20(s, 3H).

In a round-bottomed flask, 38g of a conjugated triazabicyclo [4.4.0] compound was introduced]Polymers of dec-5-ene resin (Fluka) were suspended in 160ml ethanol. Mixing 40ml of ethanolThe amino acid product of example 2lF (7.5g) was added to the stirred resin syrup. 6.5g (53mmoles) of ethyl acetimidate are added in portions to the reaction. The reaction was stirred under nitrogen for 16 hours. The resin was filtered and the filter was washed with 100ml ethanol containing 10ml concentrated HCl. The combined filtrates were concentrated to give 12g of crude title product as a pale brown viscous semi-solid. The yield was about 60-70% and the purity of the title product was shown to be 90%. The title product was further purified by reverse phase chromatography.¹H NMR(D₂O)δ4.74(s，D₂O exchange) 3, 37(t, 2H), 3.08(d, 1H), 2.93(d, 1H), 2.74(m, 2H), 2.06(s, 3H), 1.48(s, 3H).

Example 22

S- [2- [ (1-Iminoethyl) amino ] ethyl ] -2-methyl-L-cysteine, dihydrochloride salt example-22A) N-Boc-cysteamine

A3L 4-neck RB flask was purged with nitrogen for 20 minutes, and then 2-aminoethanethiol hydrochloride (113.6g, 1mol), di-tert-butyl dicarbonate (218.3g, 1mol) and 500ml of toluene were successively added. The mixture was cooled with an ice water bath and purged with nitrogen for 10 minutes. Sodium hydroxide (2.5N, 880ml, 2.2mol) was added to the stirred mixture over about 1.5 hours at 0-11 ℃. After the hydroxide addition was complete, the cold bath was removed and the resulting reaction mixture was allowed to warm to room temperature and stirred at room temperature overnight. A solution of the title compound was obtained.

example-22B)

The product solution of example-22A was cooled with an ice-water bath. A sample of chloroacetone (101.8g, 1.1mol) was added to the vigorously stirred reaction mixture over about 50 minutes at 8-11 deg.C. After the addition of the chloroacetone was complete, the cooling bath was removed and the resulting reaction mixture was stirred at room temperature overnight. The toluene layer was separated, washed with water (250ml) and concentrated in vacuo at 85 ℃ in a rotary evaporator, followed by high vacuum to give the crude title compound (225.7g, 96.7%).¹HNMR(CDCl₃，400MHz)δ4.95(bs，1H)3.20(m，4H)，2.54(t，2H)，2.20(s，3H)，1.35(s，9H)。

example-22C [2- [ [ (4-methyl-2, 5-dioxo-4-imidazolidinyl) methyl ] thio ] ethyl ] carbamic acid, 1, 1-dimethylethyl ester

To a 3L 4-neck RB flask equipped with an overhead stirrer, thermocouple and condenser coupled to an empty beaker and alkaline treater (calstic trap) were added the product of example-22B (70g, 0.3mol), absolute ethanol (80ml), sodium cyanide (19.1g, 0.39mol), ammonium carbonate (43.3g, 0.45mol) and water (720mol) in that order. The 4 th neck was plugged with a plug. The resulting reaction mixture was heated at 67-68 ℃ for 6 hours. The almost clear brown solution was then cooled to room temperature. Upon cooling, a solid began to form and the heterogeneous mixture was stirred at room temperature overnight. The reaction mixture was then acidified to pH2 with 12% hydrochloric acid at-2 to 2 ℃ for about 1 hour. The cooled reaction mixture was stirred at pH2 for an additional 30 minutes and then filtered. The flask was rinsed with distilled water (2X 250ml) and each rinse was used to wash the solid cake. The solid was washed with distilled water (2X 250ml) and air-dried for four days. The dry solid was triturated with 200ml of toluene for half an hour. The slurry was filtered. The solid was rinsed successively with toluene (50ml) and 1: 4 toluene/hexane (100ml) and air dried overnight at room temperature to give the title compound in 83.1% yield, m.p. 134-136 ℃.¹H NMR(DMSO_d6，400MHz)δ10.62(s，1H)，7.85(s，1H)，6.83(m，0.9H)，6.48(bs，0.1H)，3.29(s，2H)，2.99(m，2H)，2.71(s，2H)，2.95(m，2H)，1.32(s，9H)，1.24(s，3H)；¹³C NMR(DMSO_d6400MHz), delta 178.1, 157.1, 156.1, 78.4, 63.7, 40.7, 39.4, 33.2, 28.9, 23.8. To C₁₂H₂₁N₃O₄Analytical calculation of S: c, 47.51, H, 6.98; n, 13.85, S, 10.57. Found C, 47.76, H, 6.88, N, 13.77; s, 10.75.

example-22D) R and S- [2- [ [ (4-methyl-2, 5-dioxo-4-imidazolidinyl) methyl ] thio ] ethyl ] carbamic acid, 1, 1-dimethylethyl ester

S enantiomer R enantiomer

The reaction product of example-22C was separated into its R and S enantiomers on a Chiralpark ® AD column and eluted with methanol. The S isomer is eluted first, followed by the R enantiomer. Both isomers were used for the subsequent conversion reaction.

The S enantiomer:

[α]in MeOH at 25 ℃ +43.0(365 nm).¹H NMR：(400MHz，CD₃OD)δ1.49(s，9H)，2.05(s，3H)，2.65(t，2H)，2.9(q，2H，d)，3.20(m，2H)。IR：λcm^-1＝1772，1709。

To C₁₂H₂₁N₃O₄Analytical calculation of S (formula weight 303.38): c, 47.51, H, 6.98; and N, 13.85. Measured value: c, 47.39, H, 6.62, N, 13.83. M + H304.

The R enantiomer:

[α]in MeOH, 25 ℃ ═ 46.3(365 nm).¹H NMR：(400MHz，CD₃OD)δ1.48(s，9H)，2.05(s，3H)，2.65(t，2H)，2.85(q，2H，d)，3.18(m，2H)。IR：λcm^-1＝1770，1711。

To C₁₂H₂₁N₃O₄Analytical calculation of S (formula weight 303.38): c, 47.51, H, 6.98; and N, 13.85. Measured value: c, 48.15, H, 7.04, N, 14.37. M + H304.

example-22E) S- (2-aminoethyl) -2-methyl-L-cysteine

The acid hydrolysis method comprises the following steps:

the R-isomer product of example-22D (45, 8g, 150.9mmol) was added to a 500ml three-necked round bottom beaker equipped with a distillation condenser and treated in portions with 48% aqueous HBr (160ml) with stirring at room temperature. After the evolution of gas had ceased, the mixture was heated with a heating mantle until the flask temperature reached 126 ℃ at which time volatile tert-butyl bromide (boiling point 72-74 ℃) and thereafter a small amount of aqueous HBr (ca. 15ml) was distilled off. The distillation condenser was replaced by a reflux condenser, and the mixture was heated under reflux for 30 hours. The solution was concentrated and the residue dissolved in water (250mL) and loaded onto Dowex ® 50WX4-200 ion exchange resin (8.5X 11cm) and eluted with water (2L) followed by diluted aqueous amine hydroxide (30mL of 28-30% ammonium hydroxide diluted to 1000mL, 3L with water). Fractions containing the desired product were combined, concentrated and dried under vacuum at 75-80 ℃ for 2 hours to give 22.1g (82%) of the title product, S- (2-aminoethyl) -2-methyl-L-cysteine, as a white solid. The proton and C-13NMR spectra were consistent with the title compound. Melting point 157 ℃.¹H NMR：(400MHz，D₂O)δ1.19(3H，s)，2.53(1H，d，J＝13.6Hz)，2.57-2.72(2H，m)，2.92(1H，d，J＝13.6Hz)，2.92(2H，t，J＝6.8Hz)；¹³C NMR(100MHz，D₂O)δ24.7，31.3，38.9，40.9，59.6，180.7。

To C₆H₁₄N₂O₂S+0.1H₂Analytical calculation of O: c, 40.02, H, 7.95; n, 15.56; s, 17.81. Found C, 39.93; h, 7.98; n, 15.38; s, 17.70.

The alkaline hydrolysis method comprises the following steps:

a stainless steel autoclave equipped with a stirrer was charged with 24.2g (0.08moles) of the R-isomer product of example-22D. After purging the apparatus with nitrogen, 128g (0.32moles) of 10% caustic was added to form a solution. The autoclave was sealed and heated to 120 ℃ for 30 hours. After cooling to room temperature, the autoclave was evacuated to yield 142ml (151g) of an aqueous solution of the sodium salt of the title compound.¹H NMR: (samples were acidified with HCl and treated with D₂O dilution, 400MHz) δ 1.47(s, 3H), 2.75(m, 2H), 2.90(d, 1H, J ═ 14.8Hz), 3.06(t, 2H, J ═ 6.4Hz), 3.14(d, 1H, J ═ 14.8 Hz);¹³c NMR (samples acidified with HCl and D₂O dilution, 100MHz) δ 172.9, 60.8, 39.1, 39.0, 30.4, 22.2. MS (MS/CI-LC) M + 1179.

DBU (218L; 1.46mmol) and ethyl acetimidate hydrochloride (171 mg; 1.34mmol) were dissolved in ethanol (6ml) at room temperature (ca. 20 ℃ C.) in a 25ml single-necked round bottom flask. The title product of example-22E (200 mg; 1.12mmol) was added in one portion to the solution. The mixture was stirred until the title product of example-22E was consumed (1-2 hours). The mixture was cooled with an ice bath and treated with 6M HCl (830L).¹HNMR analysis showed a chemical yield of 95% molar or greater. The solvent was evaporated and the title product of example-22 was purified by reverse phase or ion exchange chromatography.

210g of the solution (title product of example-22E containing about 20g of basic hydrolysis reaction product) was placed in a 500ml three-necked round bottom flask. The apparatus was equipped with a mechanical stirrer, Dean-Stark apparatus (20ml with piston), condenser and temperature controller. From this mixture water (140m) was distilled off. 1-Butanol (150ml) was added to the vessel and the residual water (37ml) was azeotropically distilled. Additional 1-butanol (13ml) was removed by distillation until the vessel temperature reached 117 ℃. Mixing the powderThe alcohol slurry was cooled to room temperature and filtered through a pad of celite. The salt was washed with 1-butanol (2X 20 ml). DBU (21.8L; 146mmol) and ethyl acetimidate hydrochloride (17.1 mg; 134mmol) were dissolved in 1-butanol (40mL) at room temperature in a 500mL three-necked round bottom flask. The apparatus was equipped with a mechanical stirrer, addition funnel and temperature probe. The title product of example-22E/1-butanol solution was placed in an addition funnel and added to the ethyl acetimidate/DBU solution while maintaining the vessel temperature below 25 ℃. The mixture was stirred until the starting material was consumed (2-3 hours). A solution of concentrated HCl (94ml) and water (100ml) was placed in a 1L three-necked round bottom flask and cooled to 0 ℃. The apparatus was equipped with a mechanical stirrer, addition funnel and temperature probe. The reaction mixture was placed in an addition funnel. The reaction mixture was added to aqueous HCl while maintaining the vessel temperature below 25 ℃. Ethyl acetate (100ml) was added to the solution, and the layers were separated. The aqueous layer was washed with ethyl acetate (100ml) more than once,¹HNMR analysis showed a chemical yield of 95% molar or greater. The title product of example-22 was purified by reverse phase or ion exchange chromatography.¹HNMR(400MHz，D₂O)1.49(3H，s)，2.08(3H，s)，2.74(2H，m)，2.91(1H，d)，3.17(1H，d)，3.35(2H，t)。

Example 23

S- [2- [ (1-iminoethyl) amino ] ethyl ] -2-methyl-L-cysteine methyl ester, dihydrochloride

The product of example 22 (1.0g, 3.42mmol) dissolved in dry methanol (40ml) was charged to a 500ml three neck round bottom flask equipped with a magnetic stirrer and a thermocouple. The reaction was cooled to 0 ℃ under nitrogen. HCl gas was bubbled through the reaction for 1 minute. The reaction mixture was warmed to room temperature and stirred continuously overnight. A sample was taken from the reaction mixture and concentrated. NMR and mass spectrometry analysis showed the starting material and product. The solvent was stripped, the oily residue redissolved in dry methanol (40ml), cooled to 0 ℃, and HCl gas was again bubbled through the solution for 1 minute. The reaction mixture was warmed to room temperature and stirred overnight. A sample was taken from the reaction mixture and concentrated. NMR and mass spectrometry analysis showed the starting material and product. The solvent was stripped, the oily residue redissolved in dry methanol (40ml), cooled to 0 ℃, and HCl gas was again bubbled through the solution for 1 minute. The reaction mixture was warmed to room temperature and stirred overnight. A sample was taken from the reaction mixture and concentrated. NMR and mass spectrometry analysis showed only the desired title product. The reaction mixture was concentrated to give 1.01g of a pale yellow oil with a yield of 97%. The reaction mixture was stirred in acetonitrile (50ml) for 3 hours, and 484mg of the title product was recovered as a white fine powder. Mass spectrometry analysis: (ZMD Waters Micromass), M + H at 234.2

¹HNMR：(400MHz，D₂O)δ1.51(s，3H)，2.09(s，3H)，2.72(t，2H)，2.97(d，1H)，3.19(d，1H)，3.36(t，2H)，3.73(s，3H).¹³CNMR δ18.58，21.69，30.79，37.79，41.58，54.24，60.75，165.41，171.35。

To C₉H₁₉N₃O₂S+2HCl+0.3H₂Analytical calculation of O (311.66): c34.68, H, 6.98, N13.48, cl22.75, S10.29. Measured value: c34.51, H6.99, N13.75, cl22.75, S10.43.

Example 24

2-methyl-S- [2- (3-methyl-5-oxo-1, 2, 4-oxadiazol-4 (5H) -yl) ethyl ] -L-cysteine, monotrifluoroacetate

example-24A) N' -hydroxyethylimidoamide (ethanimide)

To a 3L round bottom flask was added hydroxylamine hydrochloride (138.98g, 2.0mol) in ethanol (1.2L) and sodium ethoxide (136.1g, 2.0mol) was added slowly. Keeping the temperature at 25-30 ℃. The reaction was stirred at room temperature for an additional 30 minutes. The precipitated NaCl was filtered off and washed with ethanol (100 ml). Propionitrile (112.75g, 2.75mol) and hydroxylamine free base in the filtrate were charged to a 3L flask. The mixture was then refluxed overnight. After cooling, the solvent was carefully removed in vacuo to 50% of its original volume. The reaction was placed in an ice bath for one hour, crystals formed and were filtered off. The filtrate was again carefully concentrated to 50% of its original volume. The reaction was placed in ice and the resulting crystals were isolated by filtration again to give 52g (35%) of the title product.

example-24B) potassium 3-methyl-1, 2, 4-oxadiazolin-5-one

A25 mL round bottom flask was charged with the product of example-24A (1g, 0.013mol), potassium tert-butoxide (1.59g, 0.013mol), and diethyl carbonate (8.18mL, 0.067 mol). The reaction was heated at reflux for 5 hours. The solvent was removed and the resulting solid was triturated with dichloromethane and diethyl ether. The solid title product was dried under high vacuum to yield 1.57g (87%).¹HNMR(d₆-DMSO，300MHz)δ1.69(bs，3H)。¹³C NMR(d₆-DMSO，99MHz)δ13.26，166.54，173.99。

example-24C) 3-methyl-1- (1-bromoethyl) -2, 4-oxadiazolin-5-one

To a 250ml round bottom flask was added the title product of example-24B, 3-methyl-1, 2, 4-oxadiazoline in DMF (100ml)Potassium 5-keto (10.13g, 0.0734 mol). To the slurry was added 1, 2-dibromoethane (31.54mL, 0.366 mol). The reaction was heated in an oil bath for 2 hours at 130 ℃. The oil bath was removed and after the reaction was cooled, water (200ml) and ethyl acetate (50ml) were added. The collected organics were washed with 3X 100ml brine. The organics were over MgSO₄Drying and concentration in vacuo gave 9.1g (60%) of the title compound.

¹HNMR(CDCl₃，300MHz)δ2.21(s，3H)，3.12(t，2H)，3.91(t，2H)。

75ml of methanol in a 100ml round bottom flask was deoxygenated by bubbling nitrogen through for 5 minutes. To 50ml of this methanol was added NaOH (1.6g, 0.040 mol). After complete dissolution of the NaOH, the suspension was stirred under an oil bath at 45 ℃ for 30 minutes. The resulting solution was cooled to room temperature and α -methyl cysteine (1.72g, 0.010mol) in 10ml of deoxygenated methanol was added. The reaction was stirred at room temperature for 45 minutes. To this reaction was added 10ml of the product of example-24C (2.07g, 0.010mol) in deoxygenated methanol. After stirring overnight the reaction was complete by mass spectrometry. The reaction mixture was diluted with water (100ml) and purified by reverse phase chromatography to give 3.0g (93%) of the title product of example 24 as its trifluoroacetate salt. M.S.M + H⁺(262.0)、M+Na⁺(282.0).¹HNMR(CD₃OD，300MHz)δ1.39(s，3H)，2.23(s，3H)，2.74(m，2H)，2.84(m，2H)，3.72(t，2H).

Example 25

[2- [ [ [ (4R) -4-methyl-2, 5-dioxo-4-imidazolidinyl ] methyl ] thio ] ethyl ] (1-N-iminoethyl) amine

The product of example-22D [2- [ [ [ (4R) -4-methyl-2, 5-dioxo-4-imidazolidinyl]Methyl radical]Thio group]Ethyl radical]Carbamic acid, 1, 1-dimethylethyl ester isomer (2.05g, 6.5mmol) dissolved in 25ml 4.0N HCl in dioxaneThe solution was stirred for 10 minutes. After addition of 2N HCl (5ml), the reaction was stirred for a further 2 hours. The reaction mixture was concentrated under reduced pressure to give 1.68g of a reddish brown binding solid. The material was dissolved in 25ml of deionized water and the pH adjusted to 8.4 with 2N NaOH. Ethyleneimidate hydrochloride (2.39g, 0.019mol) was added while maintaining the pH at 8.4. The reaction mixture was stirred at room temperature for one hour at pH 8.4. The reaction mixture was adjusted to pH 3.5 by addition of an appropriate amount of 1N HCl and stirred for a further 16 hours. The reaction mixture was concentrated on a rotary evaporator to give the crude product, which was purified by Gilson preparative HPLC to give the desired product as a white hygroscopic solid in a yield of 70%. Mass M⁺¹245 in water at 25 ℃ [ α ]]＝-37.6(365nm)。

To C₉H₁₆N₄O₂S+1.0HCl+1.3H₂Calculated and analyzed value of O (formula weight: 304.20): c35.54, H6.49, N18.42, Cl 11.65 and S10.54. Measured value: c35.83, H6.08, N18.55, Cl 11.19, S10.63.

Example 26

[2- [ [ [ (4S) -4-methyl-2, 5-dioxo-4-imidazolidinyl ] methyl ] thio ] ethyl ] (1-N-iminoethyl) amine hydrochloride

example-26A) (5S) -5- [ [ (2-aminoethyl) thio ] methyl ] -5-methyl-2, 4-imidazolidinedione, monohydrochloride

The product of example-22D, [2- [ [ [ (4S) -4-methyl-2, 5-dioxo-4-imidazolidinyl ] methyl ] thio ] ethyl ] carbamic acid, 1, 1-dimethylethyl ester isomer (2.05g, 6.5mmol) was purified by chromatography on Biotage Flash 75 silica gel with ethyl acetate in 66% toluene. A sample of this starting material (5.9g, 16.5mmol, [ α ] ═ 45.7 in MeOH at 25 ℃, 365nm) was dissolved in 165ml of thf and treated with 4.125ml of 4.0N HCl in dioxane. The reaction was stirred at room temperature for two hours and monitored by TLC. The free amine product was purified by chromatography on reversed-phase medium (YMC-ODS-AQ) to give 4.8g of the title compound.

A3.5 g (17.2mmol) sample of the product of example-26A was adjusted to a pH of 9-10 with 10% NaOH solution. To this solution was added 4.26g of ethyl acetimidate hydrochloride while adjusting the pH to 9 by adding 10% NaOH. After stirring at pH9 for 2 hours, the pH was adjusted to 7.5 by addition of the appropriate amount of 0.1N HCl. The solution was stirred for an additional 2 hours before adding 0.1N HCl to further adjust the pH to 4.5. After stirring the solution for 10 hours, the water was removed under reduced pressure (11mbar) in a water bath at 47 ℃. The crude title product was purified by chromatography on reverse phase medium (YMC-ODS-AQ) to give 156mg of the title material. Alpha in water at 25 ℃]+54.8(365 nm). To C₉H₁₆N₄O₂S+1.0HCl+0.85H₂Calculated analysis value of O (formula weight 296.09): c36.51, H6.37, N18.92, Cl 11.97, S10.83. Measured value: c36.69, H6.32, N18.85, Cl 11.46, S11.12.

Example 27

N- (ethoxycarbonyl) -S- [2- [ (1-iminoethyl) amino ] ethyl ] -2-methyl-L-cysteine, monohydrochloride

A sample of the product of example 1 (3.22g, 0.01mol) was dissolved in 50ml of deionized water, to which K was added₂CO₃(2.76g) followed by ethyl chloroformate (1.08g, 0.01 mol). The reaction mixture was stirred at 25 ℃ for 1 hour and concentrated on a rotary evaporator to give a white solid. The solid was purified by HPLC to give the desired product.

Mass M⁺¹＝292

Example 28

S- (2-aminoethyl) -2-methyl-D-cysteine dihydrochloride

[2- [ [ [ (4S) -4-methyl-2, 5-dioxo-4-imidazolidinyl) methyl ] of example-22D]Thio group]Ethyl radical]A sample of the carbamic acid, 1, 1-dimethylethyl ester product (1.025g, 3.25mmol) was dissolved in 35ml of concentrated HCl and stirred at reflux temperature for 46 hours. The reaction was concentrated under reduced pressure to give 900mg of a reddish brown binding solid. The crude product was purified by reverse phase HPLC to give pure S- (2-aminoethyl) -2-methyl-D-cysteine dihydrochloride (800mg, 98% yield). MassM⁺¹179 in water at 25 ℃ [ α]＝-85.6(365nm)。

To C₆H₁₄N₂O₂S+2HCl+1H₂O+1.6NH₄Calculated analytical values for Cl (formula weight: 356.39; accurate mass 178.07): c20.22, H7.35, N14.15, Cl 35.81, S9.00. Measured value: c20.09, H6.95, N14.55, Cl 36.15, S9.56.

Example 29

[2- (1-iminoethylamino) ethyl ] -2-methyl-D-cysteine hydrochloride

A sample of the product of example 28 (1.25g, 0.005mol) was dissolved in 20ml of deionized water and the pH was adjusted to 8.5-9 with 0.1N NaOH. The pH was maintained at 8.5 and ethyl acetimidate hydrochloride (2.39, 0.019mol) was added to the stirred reaction. The reaction was stirred at 25 ℃ and pH8.5 for 2 hours. The pH of the reaction mixture was adjusted to 4.0 by addition of the appropriate amount of 0.1N HCl. Then the reaction is carried out by a rotary evaporatorThe mixture was concentrated and the crude residue was purified by a Gilson HPLC system using a YMC AQ column using 1.1% AcOH/CH₃CN/H₂And eluting with O to obtain the desired product with quantitative yield. Mass M⁺¹Alpha in water at 220.25 ℃]＝-134.5(365nm)。

To C₈H₁₇N₃O₂S+1.2HCl+2H₂Calculated analysis of O (formula weight 299.09; accurate mass 219.10): c32.13, H7.48, N14.05, Cl 14.22, S10.72. Measured value: c32.39, H7.26, N14.05, Cl 14.33, S10.42.

Example 30

S- [2- [ (1-iminoethyl) amino ] ethyl ] -2-methyl-L-cysteine, acetic acid salt

Bio-Rad AG 1X 8 resin (300g, 960meqv.), 200-400 mesh the acetate form was slurried in HPLC grade water and loaded onto an 8cm diameter column. Before 37g (116mmol) of the product of example 1 dissolved in 10ml of water are charged to the column, the water is drained to the top of the column. The material was eluted with 1L of water. After removal of water under reduced pressure, the first 200ml portion was product free, but the subsequent 500ml gave 30g of the desired title product as a white glassy solid.

To C₈H₁₇N₃O₂S+CH₃COOH+1.3H₂Calculated analysis value of O: c39.67, H7.86, N13.88. Measured value: c39.96, H7.87, N13.69.

Example 31

S- [2- [ (1-iminoethyl) amino ] ethyl ] -2-methyl-D-cysteine, acetic acid salt

A sample of the product of example 29 (101mg, 0.33mmol) as its hydrochloride salt was converted to the title mono acetate salt by the method of example 30. To C₈H₁₇N₃O₂S.CH₃COOH+0.05HCl+2.2H₂Calculated analysis value of calculated analysis of O: c37.41, H8.01, N13.2, Cl 0.56. Measured value: c37.30, H7.92, N13.17, Cl 0.41.

Example 32

S- [2- [ (1-iminoethyl) amino ] ethyl ] -2-methyl-L-cysteine, monohydrochloride

This material was prepared by passing the product of example 1 through a reverse phase column using the conditions described for example 28.

To C₈H₁₇N₃O₂S+1.05HCl+0.8H₂Calculated analysis value of O: c35.35, H7.36, N15.44, Cl 13.65. Measured value: c35.33, H7.28, N15.45, Cl 3.68.

Example 33

D-galacturonate of S- [2- [ (1-iminoethyl) amino ] ethyl ] -2-methyl-L-cysteine

D-galacturonic acid monohydrate (0.21g, 0.001mole) was added to a stirred solution of 10ml of the 0.001M acetate product of example 30. After stirring for 2 hours, the solution was concentrated in vacuo. The title galacturonate was dissolved in 10ml water and lyophilized.

Example 34

Succinate salt of S- [2- [ (1-iminoethyl) amino ] ethyl ] -2-methyl-D-cysteine

The title material was prepared by the method of example 33 from succinic acid and the product of example 31.

To C₈H₁₇N₃O₂S+C₄H₆O₄+1.5H₂Calculated analysis value of O: c39.55, H7.19, N11.53. Measured value: c39.24, H6.04, N11.41.

Example 35

Succinate salt of S- [2- [ (1-iminoethyl) amino ] ethyl ] -2-methyl-L-cysteine

The title material was prepared by the method of example 33 from succinic acid and the product of example 30.

To C₈H₁₇N₃O₂S+C₄H₆O₄+1.1H₂Calculated analysis value of O: c39.99, H7.40, N12.33. Measured value: c40.35, H7.11, N11.76.

Example 36

Ethanolamine salt of S- [2- [ (1-iminoethyl) amino ] ethyl ] -2-methyl-L-cysteine

The title material was prepared from ethanolamine and the product of example 30 by the method of example 33.

To C₈H₁₇N₃O₂S+C₂H₇NO+2HCl+1.3H₂Calculated analysis value of O: c31.73, H7.67, N14.80, Cl 18.73. Measured value: c31.41, H7.60, N15.00, Cl 19.12.

Example 37

Ethylenediamine salt of S- [2- [ (1-iminoethyl) amino ] ethyl ] -2-methyl-L-cysteine

The title material was prepared from ethylenediamine and the product of example 30 by the method of example 33.

To C₈H₁₇N₃O₂S+2HCl+C₂H₈N₂+1.2H₂Calculated analysis value of O: c32.12, H7.92, N18.73, Cl 18.96. Measured value: c31.90, H9.19, N18.08, Cl 19.11.

Example 38

DL-aspartic acid salt of S- [2- [ (1-iminoethyl) amino ] ethyl ] -2-methyl-L-cysteine

The title material was prepared from DL-aspartic acid and the product of example 30 by the method of example 33.

To C₁₂H₂₄N₄O₆S+1.8H₂Calculated analysis value of O +0.4HOAc (formula weight 408.86): c37.60, H7.20, N13.70. Measured value: c37.59, H7.66, N13.73.

Example 39

D-glutamate salt of S- [2- [ (1-iminoethyl) amino ] ethyl ] -2-methyl-L-cysteine

The title material was prepared by the method of example 33 from D-glutamic acid and the product of example 30.

To C₁₃H₂₆N₄O₆S+1.8H₂Calculated analysis value of O +0.3HOAc (formula weight 416.88): c39.18, H7.45, N13.44. Measured value: c39.47, H7.52, N13.29.

Example 40

Citrate salt of S- [2- [ (1-iminoethyl) amino ] ethyl ] -2-methyl-L-cysteine

The title material was prepared from citric acid and the product of example 30 by the method of example 33.

To C₁₄H₂₅N₃O₉S+0.5H₂Calculated analysis of O +0.1HOAc +0.15EtOH (formula weight 433.36): c40.19, H6.35, N9.70. Measured value: c40.32, H5.74, N9.58.

EXAMPLE 41

DOWEX50WX4-400 ion exchange resin salts of S- [2- [ (1-iminoethyl) amino ] ethyl ] -2-methyl-L-cysteine

DOWEX ® 50WX4-400(3g, 1.6meq/ml, 4.8meq/g) was washed with deionized water (all water used in this experiment was deionized water) until the pH of the wash was 6. The resin was dried at room temperature for 1 hour. The product of example 30 (0.6g) in 30ml of water was charged to DOWEX resin (0.2 g). At room temperature, with ORBIT^TMThe suspension was shaken for 3 hours by a shaker. Then stripped to dryness. This procedure was repeated three times with 30ml of fresh water added after each concentration of the reaction mixture. The slurry was shaken overnight at room temperature with the final portion of fresh water.

After the reaction was stripped to dryness, 15ml of water was added. The resin was filtered and washed three more times with 15ml water. The filtrate was concentrated (a few drops of acetic acid were added) and dried in vacuo to yield 0.4g of a solution prepared from¹HNMR(D₂O) certified starting material SC-84250. The loaded resin was dried on a bench at room temperature and further dried under vacuum for 1 hour to give 0.3g of the loaded resin. A sample of this resin and a sample of unreacted washed DOWEX50WX4-400 were analyzed by nitrogen combustion: the result for the untreated resin was that the percentage (%) of N was 0%; the percentage (%) of N to the loaded resin was 9.72%.

Example 42

Potassium hydrogen sulfate salt of S- [2- [ (1-iminoethyl) amino ] ethyl ] -2-methyl-L-cysteine

By the method of example 33, from 0.001mole of KHSO₄And the product of example 30.

To C₈H₁₇N₃O₂S+KHSO₄+2H₂Calculated analysis value of O: c24.75, H5.68, N10.90, S16.00. Measured value: c24.54, H5.66, N10.73, S16.38.

Example 43

Potassium hydrogen sulfate salt of S- [2- [ (1-iminoethyl) amino ] ethyl ] -2-methyl-L-cysteine

By the method of example 33, from 0.001mole of KHSO₄And the product of example 30.

Example 44

Bisulfate salt of S- [2- [ (1-iminoethyl) amino ] ethyl ] -2-methyl-L-cysteine

To a stirred 10ml solution of the product of example 30 was added 2ml of 0.505NH₂SO₄. After stirring for 2 hours, the solution was concentrated in vacuo. The resulting salt was dissolved in 10ml of water and lyophilized.

To C₈H₁₇N₃O₂S+0.5H₂SO₄+1.5H₂Calculated analysis value of O: c32.58, H7.23, N14.75, S16.42. Measured value: c32.53, H7.17, N14.23, S16.28.

Example 45

Glycerate of S- [2- [ (1-iminoethyl) amino ] ethyl ] -2-methyl-L-cysteine

S-glyceric acid can be prepared by stirring its calcium salt with Dowex ® 50W resin in its acid form for 4 hours. Filtering the resin and reacting with H₂And O washing. The resulting filtrate was concentrated and dried in vacuo.

To C₃H₆O₄Calculated analysis value of (2): c31.31, H6.13. Measured value: c31.29, H6.19.

S-glycerate was prepared by the method described in example 33, starting from the product of example 30 with 0.001mole of S-glyceric acid.

To C₁₁H₂₃N₃O₆S+1.5H₂Calculated analysis value of O: c37.49, H7.44, N11.92, S9.10. Measured value: c37.49, H7.31, N11.73, S9.22.

Example 46

Malic acid salt of S- [2- [ (1-iminoethyl) amino ] ethyl ] -2-methyl-L-cysteine

The malate salt was prepared using the method described in example 33 starting from the product of example 30 together with 0.001mole of malic acid.

To C₈H₁₇N₃O₂S+1.33H₂O+C₄H₆O₅Calculated analysis value of (2): c38.20, H6.85, N11.15. Measured value: c38.37, H6.51, N11.09.

Example 47

Hemimalate salt of S- [2- [ (1-iminoethyl) amino ] ethyl ] -2-methyl-L-cysteine

The malate salt was prepared using the method described in example 33, starting with the product of example 30 and 0.0005mole of malic acid.

To C₈H₁₇N₃O₂S+1.75H₂O+0.5C₄H₆O₅Calculated analysis value of (2): c37.92, H7.48, N13.22. Measured value: c37.92, H7.88, N13.03.

Example 48

Potassium dihydrogen phosphate salt of S- [2- [ (1-iminoethyl) amino ] ethyl ] -2-methyl-L-cysteine

The title salt was prepared from the product of example 30 using the method described in example 33. To C₈H₁₇N₃O₂S+KH₂PO₄+2.5H₂Calculated analytical value for O +0.66 HOAc: c25.44, H6.10, N9.55. Measured value: c25.27, H5.95, N9.80.

Example 49

Sodium dihydrogen phosphate salt of S- [2- [ (1-iminoethyl) amino ] ethyl ] -2-methyl-L-cysteine

The title salt was prepared from the product of example 30 using the procedure described in example 33.

To C₈H₁₇N₃O₂S+NaH₂PO₄+2H₂Calculated analytical value for O +0.3 HOAc: c26.26, H6.20, N10.68. Measured value: c26.57, H6.25, N10.72.

Example 50

Calcium dihydrogen phosphate salt of S- [2- [ (1-iminoethyl) amino ] ethyl ] -2-methyl-L-cysteine

The title salt was prepared from the product of example 30 using the procedure described in example 33.

To C₈H₁₇N₃O₂S+2NaH₂PO₄+2H₂Calculated analysis value of O: c19.40, H5.09, N8.48. Measured value: c19.34, H5.10, N8.56.

Example 51

Calcium phosphate salt of S- [2- [ (1-iminoethyl) amino ] ethyl ] -2-methyl-L-cysteine

The title salt was prepared from the product of example 30 using the procedure described in example 33.

To C₈H₁₇N₃O₂S+Ca(H₂PO₄)₂Calculated analytical value of +0.2 HOAc: c19.96, H4.35, N8.31. Measured value: c20.14, H5.73, N8.80.

Example 52

Calcium hydrogen phosphate salt of S- [2- [ (1-iminoethyl) amino ] ethyl ] -2-methyl-L-cysteine

The title salt was prepared from the product of example 30 using the procedure described in example 33.

To C₈H₁₇N₃O₂S+CaHPO₄+2.2HCl+H₂Calculated analysis value of O: c21.18, H4.93, N9.26. Measured value: c21.20, H5.28, N9.37.

Example 53

Calcium triphosphate salt of S- [2- [ (1-iminoethyl) amino ] ethyl ] -2-methyl-L-cysteine (1.62: 1)

The title salt was prepared from the product of example 30 using the procedure described in example 33.

To C₈H₁₇N₃O₂S+Ca₃(PO₄)₂+HOAc+3H₂Calculated analysis value of O: c14.37, H2.77, N5.03. Measured value: c14.13, H3.01, N4.71.

Example 54

Calcium phosphate salt of S- [2- [ (1-iminoethyl) amino ] ethyl ] -2-methyl-L-cysteine (1: 1)

The title salt was prepared from the product of example 30 using the procedure described in example 33.

To C₈H₁₇N₃O₂S.Ca₃(PO₄)₂+2.2HCl+2H₂Calculated analysis value of O: c14.88, H3.62, N6.51. Measured value: c15.09, H3.85, N6.23.

Example 55

Bio-Rex ® 70 salt of S- [2- [ (1-iminoethyl) amino ] ethyl ] -2-methyl-L-cysteine

The neutral form of the product of example 1, Bio-Rex ® 70 salt, was prepared using the method described in example 41. The percentage (%) of N of the results for the untreated resin was 0%; the percentage (%) of N supported by the resin was 7.93%.

Example 56

IPR (amberlite) -69 salt of S- [2- [ (1-iminoethyl) amino ] ethyl ] -2-methyl-L-cysteine

Except that the resin was first treated with 1N HCl to convert it to H⁺In addition to the form, the neutral form of the IPR-69 salt of the product of example 1 was prepared using the same procedure described in example 41. The resin is polystyrene divinyl benzene sulfonic acid resin, namely the Dowex-50. It is GMP quality but covers a wider mesh size. It is lighter than Dowex color. After washing and before loading the compound, the resin is washed in H₂Slurried in O and fine particles rising to the top were decanted. 4.9g of salt was recovered from the reaction. 7.69% N (or 0.401g SC-84250/g resin).

Example 57

IPR (Amberlite) -69 salt of S- [2- [ (1-iminoethyl) amino ] ethyl ] -2-methyl-L-cysteine

The neutral form of the IPR-64 salt of the product of example 1 was prepared by the same method described in example 41 and decantation of the fines described in example 56. The resin was identical to Bio-Rex 70, except that it was GMP quality. The only difference was that after shaking overnight, the resin was shaken two more times and stripped there. 4.3g was recovered from the reaction. 6.20% N (0.346g compound/g resin).

Example 58

Preparation of monohydrochloride from dihydrochloride salt of S- [2- [ (1-iminoethyl) amino ] ethyl ] -2-methyl-L-cysteine

About 120mg of the product of example 1 are dissolved in 3ml of DMF. 1ml of propylene oxide was added and stirred. The product precipitates. Washed with diethyl ether. The product was dissolved in water and lyophilized. Table 1 shows the elemental analysis.

Example 59

Preparation of a substituted salt of S- [2- [ (1-iminoethyl) amino ] ethyl ] -2-methyl-L-cysteine hydrochloride by AgCl precipitation

The product of example 58 was dissolved in water and a stoichiometric amount of silver salt was added. The solid was filtered. The remaining solution was lyophilized. Table 2 shows the elemental analysis, where n represents the moles of water.

Example 60

Preparation of phosphate of S- [2- [ (1-iminoethyl) amino ] ethyl ] -2-methyl-L-cysteine salt by AgCl precipitation

The product of example 1 was dissolved in water, 2moles of Ag were added per mole of product of example 1₃PO₄And mixing. Filtering outSolid and freeze-dry the resulting solution. Analyzing the resultant substance and using H₃PO₄The content of phosphate was adjusted and the analysis of elements is shown in Table 3, where x represents the number of moles of phosphoric acid.

Example 61

Preparation of mixed salts from S- [2- [ (1-iminoethyl) amino ] ethyl ] -2-methyl-L-cysteine monohydrochloride

The product of example 58 was dissolved in water. A stoichiometric amount of reagent (R) was added and the elemental analysis is shown in table 4, where x represents the number of moles of R.

Example 62

Preparation of mixed salts from S- [2- [ (1-iminoethyl) amino ] ethyl ] -2-methyl-L-cysteine dihydrochloride

The product of example 1 was dissolved in water. Base was added until pH 6. Table 5 shows the elemental analysis.

Example 63

Preparation of zinc salts from dihydrochloride of S- [2- [ (1-iminoethyl) amino ] ethyl ] -2-methyl-L-cysteine

The product of example 1 was dissolved in water. Mixed with excess zinc oxide. The resulting solution was filtered and lyophilized. Table 6 shows the elemental analysis.

Example 64

Mixed salts are prepared from neutral compounds of S- [2- [ (1-iminoethyl) amino ] ethyl ] -2-methyl-L-cysteine.

The neutral form of the product of example 20 was dissolved in water. Mixed with the required reagents and lyophilized. Table 7 shows the elemental analysis, where x represents the moles of MA, metal cation and counter anion.

Example 65

Preparation of salts from neutral compounds of S- [2- [ (1-iminoethyl) amino ] ethyl ] -2-methyl-L-cysteine

Table 8 below lists the salts prepared from the product of example 1 by one of the various methods described herein and their elemental analysis. Table 8 illustrates the elemental analysis of these salts, wherein D represents a compound of formula C₈H₁₇N₃O₂S- [2- [ (1-iminoethyl) amino group of S]Ethyl radical]-2-methyl-L-cysteine, a representing the empirical formula of the acid and or source of counterions.

TABLE 1

Formula (II)	Calculated value				Measured value
										C	H	N	Cl	C	H	N	Cl
C8H17N3O2S(HCl)(H2O)	35.10	7.36	15.35	12.95	35.06	7.53	14.90	13.07

TABLE 2

TABLE 3

Formula (II)	Calculated value				Measured value
										C	H	N	P	C	H	N	P
C8H17N3O2S(H3PO4)1.5(H2O)	27.91	6.73	12.20	9.00	27.74	6.03	12.12	8.73
									C8H17N3O2S2(H3PO4)2(H2O)	21.29	6.03	9.31	13.73	20.89	5.64	9.3	13.67

TABLE 4

TABLE 5

Formula (II)	Calculated value				Measured value
												C	H	N	M	Cl	C	H	N	M	Cl
C8H16N3O2SLi2(HCl)2(H2O)	28.75	6.64	12.57	2.08	21.22	28.05	6.58	12.34	2.14	21.59
											C8H16N3O2SNa2(HCl)1.5(H2O)	28.16	6.20	12.31	-	20.78	27.71	6.10	12.47	-	22.20
C8H16N3O2SK2(HCl)2(H2O)	26.23	6.05	11.47	-	19.36	25.75	5.60	11.50	-	21.13

TABLE 6

Formula/contrast	Calculated value					Measured in fact
												C	H	N	Cl	Zn	C	H	N	Cl	Zn
C8H17N3O2S2(HCl)ZnO	25.72	5.13	11.25	18.98	17.50	25.98	4.52	11.57	20.54	17.13

TABLE 7

Formula (II)	Calculated value			Measured value
								C	H	N	C	H	N
C8H17N3O2S(NaCl)2(H2O)	30.62	6.75	13.39	30.93	6.39	13.27
							C8H17N3O2S0.5(CaCl2)1.5(H2O)	31.84	6.68	13.92	31.59	6.57	13.73

C8H17N3O2S(NaCH3SO3)2(H2O)

28.95

6.48

11.25

28.91

6.06

10.99

TABLE 8

Acid/formula	Calculated value					Measured value
												C	H	N			C	H	N
L-tartaric acid D0.6A 0.75H2OD*1.0A*0.5H2OD*1.2A*2.0H2OD*1.1A*1.5H2O	38.5938.0932.4834.37	6.916.395.755.93	12.9811.108.889.70			38.6738.0432.7634.51	6.946.435.505.81	12.6511.388.969.69
											D-tartaric acid D0.5A 0.5H2OD*1.0A*1.0H2O	39.5937.20	6.986.50	13.8510.85			39.2737.85	7.096.45	13.3410.43

(R) - (-) -mandelic acid D1.0A 2.5H2O	49.99	7.87	10.93			49.90	6.92	10.83
											(S) - (+) -mandelic acid D1.0A 2.75H2O	49.41	7.90	10.86			49.38	8.23	10.78

Citric acid D0.4A 0.8H2OD*1.0A*0.5H2O*0.1HOAc*.015EtOH	40.2240.19	7.086.35	14.239.70			40.9640.32	6.875.74	13.269.58
											Mucic acid D0.5A 2.0H2OD*0.6A*1.0H2O	36.6640.73	7.276.84	11.6612.95	S9.89		36.4638.09	7.127.13	11.2412.03	S9.30
Maleic acid D0.5A 1.5H2O	39.46	7.29	13.81			39.64	6.98	13.01
											Malonic acid D0.5A 1.5H2O	38.24	7.34	14.08			38.03	7.37	13.89

Benzoic acid D1.1A 2.75H2OD*1.0A*1.5H2OD*2.4A*1.4H2O

50.8148.9055.41

7.907.116.41

11.3211.407.82

50.5448.9355.75

6.397.456.51

11.1211.747.52

Hydrochloric acid D1.15A 2.75H2O*0.33n-PrOHD*1.0A*1.0H2OD*1.05A*0.8H2OD*1.0A*1.5H2O*0.5CaCl2D*2.0A*2.0H2O*1.0Li+D*2.0A*1.5H2O*1.0Na+D*2.0A*2.OH2O*1.0K+D*2.0A*0.5ZnOD*1.20A*1.25H2O*1.0Me4N+Cl-

36.1735.1035.3528.4128.7528.1626.2325.7236.47

8.497.367.366.266.646.206.055.138.34

14.2215.3515.4412.4212.5712.3111.4711.2514.18

13.7912.9513.6520.9621.2220.7819.3618.9819.74

Li2.08Zn17.508.11

36.2435.0635.3328.4228.0527.7125.7525.9836.44

8.107.537.286.306.586.105.604.528.66

13.9814.9015.4512.2612.3412.4711.5011.5714.18

13.5113.0713.6819.6021.5922.2021.1320.5419.84

Li2.14Zn17.136.92

Methanesulfonic acid D1.0A 1.25HCl 0.8H2OD*2.0A*0.75HCl*5.0H2OD*1.0A*1.0H2OD*1.0A-*2.0H2O*1.0Na+D*1.0A*1.0Me4N+*1.75H2O

28.8023.1032.4228.9537.17

6.406.746.956.488.52

11.198.0812.6011.2513.34

11.805.11

17.0818.5019.2315.72

28.2923.0332.0928.9137.45

6.386.587.066.067.92

11.168.1612.3610.9912.81

12.015.18

17.1718.5819.6714.95

Tosic acid D1.0A 1.5H2OD*2.2A*2.4H2OD*1.0A*1.5HCl*1.0H2OD*1.0*1.0H2OD*1.0A-*1.0Me4N+*1.0H2OD*1.0A*1.0A-*1.0Me4N+*1.0H2OD*1.2A*1.0H2OD*2.1A*5.0H2O

45.0746.3638.8143.9947.2847.8342.7438.57

6.826.556.196.657.946.796.186.22

9.356.939.0510.2611.618.589.846.55

11.46

13.1815.6613.2914.73

45.6046.7638.9043.8347.3947.9542.9438.87

6.556.016.376.757.907.255.835.75

9.216.658.939.9110.408.499.346.50

11.57

13.8014.5012.9114.77

Hydrobromic acid D1.1A 2.25H2O0.3HOAcD*0.61A*1.0H2O	30.8532.01	7.176.04	12.5514.00	Br26.2526.62	10.68	30.6631.51	6.956.33	12.2013.82	Br26.7725.71	10.16
											Ethanesulfonic acid D1.2 AD 2.1A 1.0H2O	35.4531.27	6.946.80	11.968.97			35.5831.85	6.986.62	11.538.45
D- (+) -malic acid D0.55A 0.4H2O	40.80	7.08	13.99			40.66	6.95	13.40
											1-adamantane acetic acid D0.91A 3.5H2O	53.11	9.53	9.88			53.44	8.85	10.00
1-adamantanecarboxylic acid D0.95A 3.25H2O	52.57	9.37	9.97			52.61	7.73	9.92

Yellow benzoic acid D1.0A 1.0H2O	39.20	4.57	12.70		11.63	38.80	3.94	12.35		11.84
											1R- (-) -Camphorsulfonic acid D1.0A 0.75H2O	46.04	7.51	8.95		13.66	46.12	7.44	8.72		13.66
1S- (+) -Camphorsulfonic acid D1.0A 2.0H2O	46.48	7.48	9.03		13.79	46.41	7.85	8.74		14.13
											2-mesitylenesulfonic acid D1.0A 1.25H2O	46.19	7.18	9.50		14.51	45.96	8.54	7.33		14.91
1, 5-naphthalenedisulfonic acid D1.0A 2.0H2OD*0.6A*1.250H2O	39.8640.54	5.255.90	7.3510.13		17.9217.01	39.6540.03	4.254.60	7.3310.14		18.7717.44

1, 2-ethanedisulfonic acid D0.5A 0.5H2OD*0.5A*1.5HCl*1.0H2OD*1.0A*0.8HCl*1.5H2O	33.4227.9225.79	6.546.125.80	12.9910.859.02	13.756.09	19.8316.5720.66	33.0127.8426.07	6.295.965.76	12.7511.059.58	13.766.08	18.9616.0319.97
											Sulfoacetic acid D1.1A 1.1H2O	33.42	5.89	11.69		17.84	31.61	6.04	10.92		17.35
Propane disulfonic acid D0.67A 1.0HCl 1.25H2OD*0.3A*1.6HCl*1.25H2O	28.9629.34	6.286.56	10.1211.40	8.5415.39	18.0814.53	28.7229.17	6.326.71	10.1011.50	8.9615.48	18.1214.51

L- (+) -lactic acid D1.0A 1.0H2OD*1.0A*1.0H2O*1.0HCl	40.3638.20	7.706.99	12.8312.15	10.25	9.79	40.7938.32	7.847.16	12.6012.23	10.81	9.68
											Nitric acid D1.0A 1.0H2O	31.99	6.71	18.66			32.32	5.79	18.32
Acetic acid D1.0A 1.0H2OD*1.0A*1.3H2OD*1.0A*0.05HCl*2.2H2O (S-enantiomer)	39.2039.6737.41	7.907.868.01	13.7213.8813.20	0.56		39.1939.9637.30	7.157.877.92	13.5313.6913.17	0.41
											Pamoic acid D0.5A 1.5H2O	53.17	6.41	9.54		7.28	53.65	6.38	9.92		6.92

Phosphoric acid D1.0A 1.5H2OD*2.0A*2.0H2OD*1.0A-*2.5H2O*0.66HOAc*1.0K+D*1.0A-*2.0H2O*0.33HOAc*1.0Na+D*2.0A-*0.2HOAc*1.0Ca++D*1.0A-*2.2HCl*1.0H2O*0.66HOAc*1.0Ca++D*2.0A-*3.0H2O*1.0HOAc*2.0Ca++D*2.0A-*2.2HCl*2.0H2O*0.66HOAc*3.0Ca++

27.2121.2925.4426.2619.9621.1814.3714.88

6.736.036.106.204.354.932.773.62

12.209.319.5510.688.319.265.036.51

P9.0013.73

27.7420.8925.2726.5720.1421.2014.1315.09

6.035.645.956.255.735.283.013.85

12.129.309.8010.728.809.374.716.23

P8.7313.67

Succinic acid D0.5A 0.5H2O*1.0HClD*0.5A*0.5H2O*1.0HClD*1.0A*1.5H2O (S-enantiomer)	37.0939.9939.55	6.857.407.19	12.9812.3311.53	10.95		37.2240.3539.24	6.687.116.04	13.0811.7611.41	11.45
											Sodium oxide D1.0A 2.0H2O	30.62	6.75	13.39			30.93	6.39	13.27
Calcium chloride D0.5A 1.5H2OD*0.5A*2.5H2O	31.8429.09	6.685.19	13.9212.72			31.5928.40	6.576.35	13.7310.70
											D-a-galacturonic acid D1.0A 1.75H2O	37.84	7.00	9.57		7.61	37.86	7.10	9.60		7.61

Sulfuric acid D1.0A-2.0H2O*1.0K+D*0.5A*1.5H2OD*1.0A-*2.0CN3H6+*1.5H2OD*1.5A-*.0CN3H6+*0.75H2O	24.7532.5825.9730.50	5.687.236.977.39	10.9014.7527.2523.71		16.0016.4213.8613.57	24.5432.5326.1230.94	5.667.176.446.71	10.7314.2327.3223.87		16.3816.2813.3113.23
											(S) -glyceric acid D0.5A 1.5H2O	37.49	7.44	11.92		9.10	37.49	7.31	11.73		9.22
L- (-) -malic acid D1.0A 1.33H2OD*0.5A*1.75H2O	38.2037.92	6.857.48	11.1513.22			38.3737.92	6.517.88	11.0913.03

L-ascorbic acid D1.0A 2.2H2O	38.65	6.81	9.66			38.91	6.80	9.43
											3- (N-morpholino) propane sulfonate D1.0A 2.5H2O	38.04	7.87	11.83			38.41	8.12	11.64
L-cysteic acid D0.5A 2.3H2O	30.73	6.71	13.03			30.91	7.05	12.98
											(4S) -hydroxy-L-proline D1.0A 0.3H2O	44.89	7.53	15.74			44.29	7.60	15.91
Cyclopropane-1, 1-dicarboxylic acid D1.0A 1.3H2OD*0.5A*1.5H2O	41.8840.50	6.927.45	11.2713.48			42.0240.70	6.687.10	10.8312.98

2, 2-Dimethylmalonic acid D1.0A 2.2H2OD*0.5A*1.8H2O	39.9339.68	7.587.80	10.7513.22			39.8840.10	7.377.81	10.3713.38
											Ethanolamine D0.5A 1.3H2O*2.0HCl	31.73	7.67	14.80	18.73		31.41	7.60	15.00	19.12
Ethylenediamine D0.5A 1.2H2O*2.0HCl	32.12	7.92	18.73	18.96		31.90	9.19	18.08	19.11
											D, L-aspartic acid D1.0A 1.8H2O*0.4HOAc	37.60	7.20	13.70			37.59	7.66	13.73

D-glutamic acid D1.0A 1.8H2O*0.3HOAc	39.18	7.45	13.44			39.47	7.52	13.29
											Squaric acid D1.0A 0.5H2OD*0.5A*0.75H2O	42.1041.44	5.896.78	12.2714.50		9.3711.06	42.2941.43	5.726.56	12.7314.16		9.4210.85
Fumaric acid D1.0A 2.5H2O*2.5EtOH	41.20	8.34	8.48			41.99	8.33	8.58
											1-hydroxy-2-naphthoic acid D1.2A 1.0H2O*1.0EtOH	54.72	6.85	8.25			54.32	6.08	8.31

1-hydroxy-2-naphthalenesulfonic acid D0.65A 2.4H2O	43.42	6.58	10.48			51.01	6.53	10.39
											2-Carboxyethylphosphonic acid D1.1A 1.25H2O*1.0CaCl2D*1.5A*1.0H2OD*1.5A*0.75H2O*1.0LiCl	29.0833.1529.65	5.876.495.77	9.009.648.30			29.3433.1729.66	5.836.565.71	8.709.288.84
Phosphonoacetic acid D0.5A 2.5H2OD*2.0A*1.0H2O	32.5827.86	6.836.12	12.668.12			32.1627.76	6.735.56	13.338.40
											Phenylphosphonic acid D1.0A 0.5H2O	43.52	6.52	10.87			43.90	6.78	10.12

L-pyroglutamic acid D1.0A 1.2H2O	42.20	7.19	15.15			42.35	7.10	15.01
											HPF6D*1.0A*0.5H2O	26.31	4.97	11.23			26.63	5.10	10.64

Biological data

Some or all of the following tests were used to demonstrate the nitric oxide synthase inhibitory activity of the compounds of the present invention and to demonstrate useful pharmacological properties.

Nitric oxide synthase citrulline assay

Can be monitored by monitoring L2, 3-³H]Arginine to L2, 3-³H]Citrulline conversion assay Nitric Oxide Synthase (NOS) activity (Bredt and Snyder,Proc.Natl.Acad.Sci.U.S.A.， 87682-685, 1990 and Moore et al,J，Med.Chem.，39,669-672, 1996). Human inducible NOS (hinOS), human endothelial constitutive NOS (hecNOS), and human neuronal constitutive NOS (hncNOS) were each cloned from RNA extracted from human tissues. cDNA for Human Inducible NOS (HiNOS) was isolated from a lambda cDNA library prepared from RNA extracted from colon samples of patients with ulcerative colitis. cDNA of human endothelial constitutive NOS (hecNOS) was isolated from a lambda cDNA library prepared from RNA extracted from Human Umbilical Vein Endothelial Cells (HUVEC), and cDNA of human neuronal constitutive NOS (hncNOS) was isolated from a lambda cDNA library prepared from RNA extracted from human cerebellum obtained from a cadaver. The recombinant enzyme was expressed in Sf9 insect cells using baculovirus vectors (Rodi et al, in The Biology of Nitric Oxide, Pt 4: Enzymology, Biochemistry and Immunology (Biology of Nitric Oxide, Pt 4: Enzymology, Biochemistry and Immunology); Moncada, S., Feelisch, M., Busse, R., Higgs, E., eds., Portland Press Ltd.: London, 1995; pages 447 and 450). The enzyme activity was separated from the soluble cell extract and partially purified by DEAE-Sepharose chromatography. To measure NOS activity, 10. mu.L of enzyme was added to 40. mu.L of 50mM Tris (pH 7.6) in the presence or absence of test compound by adding bovine serum albumin containing 50mM Tris (pH 7.6), 2.0mg/mL, 2.0mM DTT, 4.0mM CaCl₂20 μ MFAD, 100 μ M tetrahydrobiopterin, 0.4mM NADPH and L- [2, 3-³H]Arginine 60. mu. M L-arginine 50. mu.L of the reaction mixture started the reaction. The final concentration of L-arginine in this test was 30. mu.M. For hec NOS or hnc NOS, calmodulin was included at a final concentration of 40-100 nM. Following incubation at 37 ℃ for 15 minutes, the reaction was terminated by adding 400. mu.L of a suspension of Dowex50W X-8 cation exchange resin (1 part resin, 3 parts buffer) in stop buffer containing 10mM EGTA, 100mM HEPES, pH5.5 and 1mM L-citrulline. The resin is precipitated after mixing and an aliquot of the supernatant is counted using a liquid scintillation counter to determine L2, 3-³H]-formation of glutamic acid. Results as IC of Compounds on hinOS, hecNOS and hncNOS₅₀The values are reported in table 1.

In vivo assay

Rats may be treated with intraperitoneal injections of 1-12.5mg/kg endotoxin (LPS) with or without oral administration of nitric oxide synthase inhibitors. Plasma nitrite/nitrate levels were determined 5 hours after injection. This result can be used to show that administration of a nitric oxide synthase inhibitor reduces the rise in plasma nitrite/nitrate levels, a reliable indicator of endotoxin-induced nitric oxide production.

Nitrite assay in Raw cells

RAW 264.7 cells were seeded in 96-well tissue culture plates in the presence of LPS and allowed to grow overnight (17 hours) to confluence to induce NOS. Leave an array of 3-6 wells untreated and serve as a control for nonspecific background reduction. The medium was removed from each well and the wells were washed twice with Kreb-Ringers-Hepes (25mM, pH7.4) containing 2mg/ml glucose. Then will be thinCells were placed on ice and incubated with 50. mu.L of buffer containing L-arginine (30. mu.M) +/-inhibitor for 1 hour. The test can be started by heating the plate to 37 ℃ for 1 hour in a water bath. Nitrite production by intracellular iNOS will be linear with time. The plate may be placed on ice to terminate the cell assay, and the buffer containing nitrite removed for analysis by the previously disclosed fluorometric method. T.P.Misko et al,Analytical Biochemistry，214，11-16(1993)。

human cartilage explant assay

The bone pieces were rinsed twice with Dulbecco's phosphate buffered saline (GibcoBRL) and once with Dulbecco's Modified Eagles Medium (GibcoBRL) and placed in Petri dishes with Minimal Essential Medium (MEM) without phenol Red (GibcoBRL). Cartilage was cut into small explants weighing about 15-45mg, and 1-2 explants per well were placed in 96 or 48 well culture plates with 200-500. mu.L of medium per well. The medium is a conventional modified medium with minimal essential medium (Eagle) other than L-arginine, non-L-glutamine and non-phenol red-prepared Earle's salts, or a serum-free Neuman and Tytel (GibcoBRL) conventional modified medium other than L-arginine, non-insulin, non-ascorbic acid, non-L-glutamine and non-phenol red-prepared. Both were supplemented with 100. mu. M L-arginine (Sigma), 2mM L-glutamine, 1 XHL-1 supplement (Bio Whittaker), 50mg/ml ascorbic acid (Sigma) and 150pg/ml recombinant human IL-1. beta. (RD system) prior to use to induce nitric oxide synthase. The compound was added in 10. mu.L aliquots with 5% CO₂Explants were cultured at 37 ℃ for 18-24 hours. The old supernatant was then discarded, replaced with fresh medium containing recombinant human IL-1. beta. and compound, and incubated for an additional 20-24 hours. The nitrite in the supernatant was analyzed by fluorescence (Misko et al, anal. biochem., 214, 11-16, 1993). All samples were performed in quadruplicate. Unstimulated controls were cultured in medium without recombinant human IL-1 β. IC was determined by plotting the percent inhibition of nitrite production against six different concentrations of inhibitor₅₀Values (table 1).

Table 9 shows examples of biological activities of some of the compounds of the invention.

TABLE 9 biological Activity. Values represent the average of all experiments and all study batches.

Examples of the Compounds	HiNOS IC50(μM)	hecNOS IC50(μM)	hncNOS IC50(μM)	Human cartilage IC50(μM)
					Example 1	3.1	77	15	0.7
Example 2	4.4	302	58	8.2
					Example 3	74	266	86
Example 4	197	1100	539
					Example 7	3.4	78	17
Example 11	0.9	26	6.0
					Example 16	7.2	＞100	36	0.7
Example 18	12	＞100	181
					Example 19	12	1080	220

One skilled in the art can easily ascertain the essential characteristics of this invention from the foregoing description, and without departing from the spirit and scope thereof, can make various changes and modifications of the invention to adapt it to various usages and conditions.

Claims (39)

1. S-2- [ (1-iminoethyl) amino ] ethyl ] -2-methyl-L-cysteine, or a pharmaceutically acceptable salt thereof.
2. 2. The compound of claim 1, wherein said compound is in the form of a pharmaceutically acceptable salt.
3. 3. The pharmaceutically acceptable salt of claim 2 having at least one anionic counterion.
4. 4. The pharmaceutically-acceptable salt of claim 3 wherein the anionic counterion is selected from the group consisting of halide, carboxylate, sulfonate, sulfate, phosphate, phosphonate, resin-bound anion, oxide and nitrate.
5. 5. The pharmaceutically-acceptable salt of claim 4 wherein the anionic counterion is a halide.
6. 6. The pharmaceutically acceptable salt of claim 4 wherein the halide is chloride.
7. 7. The pharmaceutically-acceptable salt of claim 4 wherein the anionic counterion is a carboxylate.
8. 8. The pharmaceutically acceptable salt of claim 7, wherein the carboxylic acid salt is selected from the group consisting of formate, acetate, propionate, trifluoroacetate, succinate, salicylate, DL-aspartate, D-aspartate, L-aspartate, DL-glutamate, D-glutamate, L-glutamate, glycerate, succinate, stearate, DL-tartrate, D-tartrate, L-tartrate, (±) mandelate, (R) - (-) mandelate, (S) - (+) -mandelate, citrate, mucate, maleate, malonate, benzoate, DL-malate, D-malate, L-malate, hemi-malate, 1-adamantane acetate, salicylate, stearate, DL-tartrate, L-tartrate, D-tartrate, L-tartrate, 1-adamantane carboxylate, xanthate, sulfoacetate, (+ -) -lactate, L- (+) -lactate, D- (-) -lactate, pamoate, D-alpha-galacturonate, glycerate, DL-cystine, D-cystine, L-cystine, DL-homocystine, D-homocystine, L-homocystine, DL-cysteine, D-cysteine, L-cysteine, (4S) -hydroxy-L-proline, cyclopropane-1, 1-dicarboxylate, 2-dimethylmalonate, squarate, tyrosine anion, proline anion, fumarate, 1-hydroxy-2 naphthoate, tyrosine anion, and mixtures thereof, Phosphonoacetate, carbonate, bicarbonate, 3-phosphonopropionate, DL-pyroglutamate, D-pyroglutamate and L-pyroglutamate.
9. 9. The pharmaceutically-acceptable salt of claim 4 wherein the anionic counterion is a sulfonate salt.
10. 10. The pharmaceutically acceptable salt of claim 9, wherein the sulfonate salt is selected from the group consisting of methanesulfonate, toluenesulfonate, benzenesulfonate, trifluoromethanesulfonate, ethanesulfonate, (±) -camphorsulfonate, naphthalenesulfonate, 1R- (-) -camphorsulfonate, 1S- (+) -camphorsulfonate, 2-mesitylsulfonate, 1, 5-naphthalenedisulfonate, 1, 2-ethanedisulfonate, 1, 3-propanedisulfonate, 3- (N-morpholino) propanesulfonate, biphenylsulfonate, isethionate, and 1-hydroxy-2-naphthalenesulfonate.
11. 11. The pharmaceutically-acceptable salt of claim 4 wherein the anionic counterion is a sulfate.
12. 12. The pharmaceutically acceptable salt of claim 4 wherein the sulfate salt is selected from the group consisting of sulfate, monopotassium sulfate, monosodium sulfate, and bisulfate.
13. 13. The pharmaceutically-acceptable salt of claim 4 wherein the anionic counterion is a phosphate.
14. 14. The pharmaceutically acceptable salt of claim 4, wherein the phosphate salt is selected from the group consisting of phosphate salts, dihydrogen phosphate salts, potassium hydrogen phosphate salts, dipotassium phosphate salts, potassium phosphate salts, sodium hydrogen phosphate salts, disodium phosphate salts, sodium phosphate salts, calcium dihydrogen phosphate salts, calcium hydrogen phosphate salts, calcium triphosphate salts, and hexafluorophosphate salts.
15. 15. The pharmaceutically-acceptable salt of claim 4 wherein the anionic counterion is a phosphonate.
16. 16. The pharmaceutically acceptable salt of claim 4 wherein the phosphonate is selected from the group consisting of vinyl phosphonate, 2-carboxyethyl phosphonate and phenyl phosphonate.
17. 17. The pharmaceutically-acceptable salt of claim 4 wherein the anionic counterion is a resin-bound anion.
18. 18. The pharmaceutically-acceptable salt of claim 4 wherein the resin-bound anion is selected from the group consisting of polyacrylate resins and sulfonated poly (styrene divinylbenzene) resins.
19. 19. The pharmaceutically-acceptable salt of claim 18 wherein the resin-bound anion is a polyacrylate-containing resin.
20. 20. The pharmaceutically-acceptable salt of claim 19 wherein the resin-bound anion is a resin comprising a polyacrylate salt crosslinked with divinylbenzene.
21. 21. The pharmaceutically-acceptable salt of claim 18 wherein the resin-bound anion is a sulfonated poly (styrene divinylbenzene) -containing resin.
22. 22. The pharmaceutically-acceptable salt of claim 21 wherein the resin-bound anion is a resin comprising sulfonated poly (styrene divinylbenzene) crosslinked with divinylbenzene.
23. 23. The pharmaceutically-acceptable salt of claim 4 wherein the anionic counterion is nitrate.
24. 24. The pharmaceutically-acceptable salt of claim 4 wherein the salt contains at least one cationic counterion.
25. 25. The pharmaceutically-acceptable salt of claim 24 wherein the cationic counterion is selected from the group consisting of an ammonium cation, an alkali metal cation, an alkaline earth metal cation, a transition metal cation, and a resin-bound cation.
26. 26. The pharmaceutically-acceptable salt of claim 25 wherein the cationic counterion is an ammonium cation.
27. 27. The pharmaceutically-acceptable salt of claim 26 wherein the ammonium cation is selected from the group consisting of ammonium, methylammonium, dimethylammonium, trimethylammonium, tetramethylammonium, ethanolammonium, dicyclohexylammonium, guanidinium, and ethylenediammonium cations.
28. 28. The pharmaceutically-acceptable salt of claim 25 wherein the cationic counterion is an alkali metal cation.
29. 29. The pharmaceutically acceptable salt of claim 28 wherein the alkali metal cation is selected from the group consisting of lithium cations, sodium cations, potassium cations, and cesium cations.
30. 30. The pharmaceutically-acceptable salt of claim 25 wherein the cationic counterion is an alkaline earth metal cation.
31. 31. The pharmaceutically acceptable salt of claim 30 wherein the alkaline earth metal cation is selected from the group consisting of beryllium cation, magnesium cation, and calcium cation.
32. 32. The pharmaceutically-acceptable salt of claim 25 wherein the cationic counterion is a transition metal cation.
33. 33. The pharmaceutically-acceptable salt of claim 32 wherein the transition metal cation is a zinc cation.
34. 34. The pharmaceutically-acceptable salt of claim 25 wherein the cationic counterion is a resin-bound cation.
35. 35. The pharmaceutically-acceptable salt of claim 34 wherein the resin-bound cation is a cationically-functionalized poly (styrene divinylbenzene) resin.
36. 36. The pharmaceutically-acceptable salt of claim 34 wherein the resin-bound cation is an aminated poly (styrene divinylbenzene) resin.
37. 37. The pharmaceutically-acceptable salt of claim 34 wherein the resin-bound cation is a cationically-functionalized polyacrylic resin.
38. 38. The pharmaceutically-acceptable salt of claim 34 wherein the resin-bound cation is an aminated polyacrylic resin.
39. 39. The compound of claim 1, wherein said compound is S- [2- [ (1-iminoethyl) amino ] ethyl ] -2-methyl-L-cysteine.