WO2017198179A1 - 新型噻唑类衍生物在治疗炎性肠病中的应用 - Google Patents

新型噻唑类衍生物在治疗炎性肠病中的应用 Download PDF

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WO2017198179A1
WO2017198179A1 PCT/CN2017/084749 CN2017084749W WO2017198179A1 WO 2017198179 A1 WO2017198179 A1 WO 2017198179A1 CN 2017084749 W CN2017084749 W CN 2017084749W WO 2017198179 A1 WO2017198179 A1 WO 2017198179A1
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group
dmso
nmr
ppm
compound
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PCT/CN2017/084749
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French (fr)
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李洪林
赵振江
王蕊
徐玉芳
王婉琦
杨迎芳
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华东理工大学
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Priority to US16/302,574 priority Critical patent/US11534427B2/en
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    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/33—Heterocyclic compounds
    • A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
    • A61K31/41—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having five-membered rings with two or more ring hetero atoms, at least one of which being nitrogen, e.g. tetrazole
    • A61K31/425—Thiazoles
    • A61K31/426—1,3-Thiazoles
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P1/00—Drugs for disorders of the alimentary tract or the digestive system
    • A61P1/04—Drugs for disorders of the alimentary tract or the digestive system for ulcers, gastritis or reflux esophagitis, e.g. antacids, inhibitors of acid secretion, mucosal protectants

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  • the present invention relates to the field of medicinal chemistry; in particular, the present invention relates to the use of novel thiazole derivatives for the preparation of a medicament for the treatment of ulcerative colitis and for the treatment of inflammatory bowel disease, in particular ulcerative colitis.
  • Ulcerative colitis also known as chronic non-specific ulcerative colitis, is an unexplained chronic inflammation of the intestinal mucosa and ulcerative lesions, involving the rectal mucosa, sigmoid mucosa, but also retrograde To the left colon, right colon, and even the entire colon and terminal ileum, and Crohn disease (CD) is called inflammatory bowel disease (IBD). Its clinical manifestations are mainly diarrhea, mucus pus and bloody stools, abdominal pain. The severity of the illness varies, and it is often a chronic course of recurrent episodes. The disease can occur at any age, more common in 20-40 years old, and can also be seen in children or old age. There was no significant difference in the incidence of men and women.
  • Pathological features are diffuse inflammation of the mucosa, infiltration of inflammatory cells such as diffuse lymphocytes, plasma cells, and monocytes in the lamina propria, and a large number of neutrophils and eosinophils infiltrating during the active period, which can occur in cryptitis and recession.
  • clinicians have an understanding of ulcerative colitis. Most researchers believe that ulcerative colitis is caused by multi-factor interactions, including infection, immunity, genetics, environment and psychology. However, to date, the specific pathogenic factors and pathogenesis of the disease have not been clarified. There has been no significant progress in the treatment of the treatment for many years, and the efficacy is not satisfactory.
  • ulcerative colitis is considered to be a disease that seriously affects people's quality of life and is extremely difficult to cure.
  • drugs for ulcerative colitis mainly aminosalicylic acid, sulfasalazine, oxarazin, mesalazine, immunosuppressive agents and hormonal drugs.
  • these drugs have a certain therapeutic effect, they also have serious side effects. Therefore, ulcerative colitis is considered to be a disease that seriously affects people's quality of life and is extremely difficult to cure.
  • the development of special effects drugs is valued by pharmaceutical companies worldwide.
  • the present invention provides the use of a compound of Formula I, or a pharmaceutically acceptable salt or ester thereof, for the manufacture of a medicament for the treatment of inflammatory bowel disease:
  • R 1 is selected from the group consisting of: H, a substituted or unsubstituted C1-C6 alkyl group, a C3-C6 cycloalkyl group;
  • R 2 is independently selected from the group consisting of: H, halogen, substituted or unsubstituted C1-C6 alkyl, substituted or unsubstituted C1-C6 alkoxy, CN, NO 2 , hydroxyl, NR a R b ;
  • R a and R b may be independently selected from H or C1-C6 alkyl
  • R 3 is selected from the group consisting of: H, a substituted or unsubstituted C1-C6 alkyl group;
  • R 4 is selected from the group consisting of: H, halogen
  • n is an integer from 0 to 4.
  • n is an integer of 0-5.
  • the inflammatory bowel disease is ulcerative colitis or Crohn's disease; preferably ulcerative colitis.
  • R 1 is selected from the group consisting of: H, substituted or unsubstituted C1-C6 alkyl
  • R 2 is selected from the group consisting of: H, halogen, substituted or unsubstituted C1-C6 alkyl, substituted Or unsubstituted C1-C6 alkoxy
  • R 3 is selected from: H, substituted or unsubstituted C1-C6 alkyl
  • R 4 is selected from: H, halogen
  • m is an integer from 0 to 2
  • n is 0 to An integer of 2.
  • the compound is as shown in Formula II:
  • R 1 is selected from the group consisting of: H, a substituted or unsubstituted C1-C3 alkyl group;
  • R 5 and R 6 are independently selected from the group consisting of: H, halogen, substituted or unsubstituted C1-C3 alkyl;
  • R 3 is selected from the group consisting of: H, a substituted or unsubstituted C1-C3 alkyl group;
  • R 7 , R 8 and R 9 are independently selected from the group consisting of: H, halogen.
  • the compound of Formula I, or a pharmaceutically acceptable salt or ester thereof is selected from the group consisting of a compound or a pharmaceutically acceptable salt or ester thereof:
  • the compound is a compound selected from the group consisting of:
  • the compound is a compound selected from the group consisting of:
  • the present invention provides the use of a pharmaceutical composition
  • a pharmaceutical composition comprising a compound of the first aspect of the invention or a pharmaceutically acceptable compound thereof for the manufacture of a medicament for the treatment of inflammatory bowel disease A salt or ester, and a pharmaceutically acceptable carrier or excipient.
  • the inflammatory bowel disease includes, but is not limited to, ulcerative colitis or Crohn's disease; preferably ulcerative colitis.
  • the present invention provides a pharmaceutical composition for treating inflammatory bowel disease and comprising the compound of the first aspect of the invention or a pharmaceutically acceptable salt or ester thereof, and a pharmaceutically acceptable An acceptable carrier or excipient.
  • the inflammatory bowel disease includes, but is not limited to, ulcerative colitis or Crohn's disease; preferably ulcerative colitis.
  • the pharmaceutical composition is a dosage form suitable for oral administration, including but not limited to tablets, solutions, suspensions, capsules, granules, powders.
  • the present invention provides a method of treating inflammatory bowel disease, the method comprising administering a compound of the first aspect of the invention or the pharmaceutical composition of the third aspect of the invention to a inflammatory bowel The object of illness.
  • the inflammatory bowel disease includes, but is not limited to, ulcerative colitis or Crohn's disease; preferably ulcerative colitis.
  • Fig. 1 shows the gross morphological damage of the colon and the pathological observation of colon tissue in the normal control group, the model group, the drug treatment group, and the positive control group (the application effect diagram of the compound 1).
  • FIG. 1 Effect of Compound 16 (312 in the figure) on the general condition of TNBS-mediated UC model mice (weights of mice were weighed daily during treatment of BALB/c mice, data are expressed as mean ⁇ SEM, #P ⁇ 0.05,##P ⁇ 0.01,###P ⁇ 0.001, compared with the normal control group. *P ⁇ 0.05, **P ⁇ , compared with the model group.)
  • Normal control mice observe stool There were no abnormalities such as diarrhea or blood in the stool; the stool characteristics of mice modeled by TNBS enema had different degrees of changes: such as loose stools or bloody stools. From the third day of the experiment, the BALB/c group except the normal control group was small.
  • the diarrhea of the control group administered with sulfasalazine was mild, only in the experiment.
  • the diarrhea status of the mice in the three different dose groups of Compound 16 was relieved compared with the model group .
  • FIG. 3 Effect of Compound 16 (312 in the figure) on colon mass in TNBS-mediated UC model mice.
  • Panel A shows representative colon photographs from different experimental groups.
  • Panel B shows the ratio of colon weight to length in each experimental group. #p ⁇ 0.05, compared with the control group. *p ⁇ 0.05, **p ⁇ 0.01, compared to the model group. It can be seen from the figure that after the establishment of the UC model, the mice in the model group developed severe lesions in the colon.
  • Different doses of compound 312 (5 mg/kg, 20 mg/kg, 50 mg/kg) and positive drugs can improve the degree of colonic ulcer lesions to varying degrees.
  • the condition of visceral adhesion, and 312 of 5 mg/kg had the best effect on the improvement of colonic lesions.
  • FIG. 4 Effect of Compound 16 (312 in the figure) on colonic histopathology in TNBS-induced UC model mice.
  • the colon tissue integrity of the model group was severely damaged and local erosion occurred.
  • Give positive drug SASP treatment After the treatment, the above lesions were largely reversed; after the administration of Compound 16, the mucosal integrity was improved to a large extent compared with the model group, and the dose of 5 mg/kg and 50 mg/kg was given at 20 mg/kg.
  • Compound 16 is more effective, with a low dose of 5 mg/kg being similar to a positive drug.
  • alkyl refers to a saturated branched or straight-chain alkyl group having a carbon chain length of from 1 to 10 carbon atoms, and preferred alkyl groups include from 2 to 8 carbon atoms, from 1 to 6, and from 1 to An alkyl group having 4 carbon atoms and 1-3 carbon atoms.
  • alkyl groups include, but are not limited to, methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, heptyl, and the like.
  • the alkyl group may be substituted by one or more substituents, for example by halogen or haloalkyl.
  • the alkyl group may be an alkyl group substituted with 1 to 4 fluorine atoms, such as a trifluoromethyl group, or the alkyl group may be an alkyl group substituted with a fluoroalkyl group.
  • cycloalkyl means a saturated alkyl group having an alicyclic structure, for example, a C3-C6 cycloalkyl group.
  • the cycloalkyl group includes, but is not limited to, cyclopropyl, cyclobutyl, cyclopentyl or cyclohexyl.
  • the cycloalkyl groups described herein may be substituted or unsubstituted, including but not limited to, substituted with one or more halogen atoms, such as a fluorine atom.
  • amino refers to a group of the formula "NR x R y ", wherein R x and R y may be independently selected from H or C 1 -C 3 alkyl or C 1 -C 3 haloalkyl. In specific embodiments herein described in the “amino” refers to NH 2.
  • halogen refers to fluoro, chloro, bromo and iodo.
  • the halogen is chlorine or fluorine; more preferably fluorine.
  • the present inventors have unexpectedly discovered a series of novel thiazole derivatives having a structural skeleton which is excellent in the treatment of inflammatory bowel diseases, particularly ulcerative colitis. These compounds show significant ulcerative colitis therapeutic activity in animal experiments.
  • the compound of the invention is a compound of formula I or a pharmaceutically acceptable salt or ester thereof:
  • R 1 is selected from the group consisting of: H, a substituted or unsubstituted C1-C6 alkyl group, a C3-C6 cycloalkyl group;
  • R 2 is independently selected from the group consisting of: H, halogen, substituted or unsubstituted C1-C6 alkyl, substituted or unsubstituted C1-C6 alkoxy, CN, NO 2 , hydroxyl, NR a R b ;
  • R a and R b may be independently selected from H or C1-C6 alkyl
  • R 3 is selected from the group consisting of: H, a substituted or unsubstituted C1-C6 alkyl group;
  • R 4 is selected from the group consisting of: H, halogen
  • n is an integer from 0 to 4.
  • n is an integer of 0-5.
  • R 1 is selected from the group consisting of: H, a substituted or unsubstituted C1-C6 alkyl group
  • R 2 is selected from the group consisting of: H, halogen (preferably F), substituted (preferably F-substituted) or unsubstituted C1 -C6 alkyl (preferably C1-C3 alkyl), substituted or unsubstituted C1-C6 alkoxy
  • R 3 is selected from: H, substituted or unsubstituted C1-C6 alkyl (preferably C1-C3 alkyl)
  • R 4 is selected from the group consisting of: H, halogen (preferably Cl); m is an integer of 0 to 2; and n is an integer of 0 to 2.
  • the compound of the invention is as shown in formula II:
  • R 1 is selected from the group consisting of: H, a substituted or unsubstituted C1-C3 alkyl group;
  • R 5 and R 6 are independently selected from: H, halogen, substituted (preferably F substituted) or unsubstituted C1-C3 alkyl (preferably methyl);
  • R 3 is selected from the group consisting of: H, a substituted or unsubstituted C1-C3 alkyl group;
  • R 7 , R 8 and R 9 are independently selected from the group consisting of H, halogen (preferably Cl).
  • the present invention further provides a pharmaceutical composition
  • a pharmaceutical composition comprising the treatment An effective amount of a compound of the invention, or a pharmaceutically acceptable salt or ester thereof, and a pharmaceutically acceptable carrier or excipient.
  • Examples of pharmaceutically acceptable salts of the compounds of the invention include, but are not limited to, inorganic and organic acid salts such as the hydrochloride, hydrobromide, sulfate, citrate, lactate, tartrate, maleate salts. , fumarate, mandelate and oxalate; and inorganic and formed with bases such as sodium hydroxy, tris(hydroxymethyl)aminomethane (TRIS, tromethamine) and N-methyl glucosamine Organic base salt.
  • inorganic and organic acid salts such as the hydrochloride, hydrobromide, sulfate, citrate, lactate, tartrate, maleate salts. , fumarate, mandelate and oxalate
  • bases such as sodium hydroxy, tris(hydroxymethyl)aminomethane (TRIS, tromethamine) and N-methyl glucosamine Organic base salt.
  • a pharmaceutically acceptable ester of a compound of the invention is a compound wherein the carboxyl group of the compound of formula I is esterified, including but not limited to, a substituted or unsubstituted C1-C10 alkyl or cycloalkyl ester of a compound of formula I, Group of esters, An ester of a group, wherein the substitution is substituted with a group selected from the group consisting of phenyl, halogen.
  • a pharmaceutically acceptable ester of a compound of the invention is selected from the group consisting of
  • the compound of the present invention or a pharmaceutically acceptable salt thereof is orally administered to a mammal daily in an amount of from about 0.0025 to 50 mg/kg body weight. Preferably, however, it is about 0.01 to 10 mg per kilogram of oral administration.
  • a unit oral dose can include from about 0.01 to 50 mg, preferably from about 0.1 to 10 mg, of a compound of the invention.
  • the unit dose may be administered one or more times per day in one or more tablets, each tablet containing from about 0.1 to 50 mg, conveniently from about 0.25 to 10 mg of the compound of the invention or a solvate thereof.
  • the pharmaceutical composition of the present invention can be formulated into a form suitable for various administration routes, including but not limited to being formulated for parenteral, subcutaneous, intravenous, intramuscular, intraperitoneal, transdermal, oral, intrathecal, intracranial A form of administration, intranasal or topical, for the treatment of tumors and other diseases.
  • the amount administered is an amount effective to ameliorate or eliminate one or more conditions.
  • an effective amount is an amount sufficient to ameliorate or in some way alleviate the symptoms associated with the disease.
  • Such doses can be administered as a single dose or can be administered according to an effective therapeutic regimen.
  • the amount administered may cure the disease, but administration is usually to improve the symptoms of the disease. Repeated administration is generally required to achieve the desired improvement in symptoms.
  • the dosage of the drug will be determined by the age of the patient, the health and weight, the type of concurrent treatment, the frequency of treatment, and the desired therapeutic benefit.
  • the pharmaceutical preparation of the present invention can be administered to any mammal as long as they can obtain the therapeutic effect of the compound of the present invention.
  • the most important of these mammals is humans.
  • the compounds of the invention or pharmaceutical compositions thereof are useful for the treatment of ulcerative colitis.
  • the pharmaceutical preparations of the invention can be made in a known manner. For example, it is manufactured by a conventional mixing, granulating, tableting, dissolving, or freeze drying process. In the manufacture of oral formulations, the mixture can be selectively milled by combining the solid adjuvant with the active compound. If necessary or necessary, after adding an appropriate amount of auxiliary agent, the mixture of particles is processed to obtain a tablet or tablet core.
  • Suitable excipients are, in particular, fillers, such as sugars such as lactose or sucrose, mannitol or sorbitol; cellulose preparations or calcium phosphates, such as tricalcium phosphate or calcium hydrogen phosphate; and binders, such as starch pastes, including corn starch. , wheat starch, rice starch, potato starch, gelatin, tragacanth, methyl cellulose, hydroxypropyl methyl cellulose, sodium carboxymethyl cellulose, or polyvinylpyrrolidone.
  • fillers such as sugars such as lactose or sucrose, mannitol or sorbitol
  • cellulose preparations or calcium phosphates such as tricalcium phosphate or calcium hydrogen phosphate
  • binders such as starch pastes, including corn starch. , wheat starch, rice starch, potato starch, gelatin, tragacanth, methyl cellulose, hydroxypropyl methyl cellulose, sodium carboxymethyl
  • a disintegrating agent such as the above-mentioned starch, and carboxymethyl starch, crosslinked polyvinylpyrrolidone, agar, or alginic acid or a salt thereof such as sodium alginate may be added.
  • Adjuvants are especially flow regulators and lubricants, for example, silica, talc, stearates such as calcium magnesium stearate, stearic acid or polyethylene glycol.
  • the tablet core can be provided with a suitable coating that is resistant to gastric juice. For this purpose, a concentrated sugar solution can be applied.
  • Such solutions may contain gum arabic, talc, polyvinylpyrrolidone, polyethylene glycol and/or titanium dioxide, lacquer solutions and suitable organic solvents or solvent mixtures.
  • a suitable cellulose solution such as cellulose acetate phthalic acid or hydroxypropyl methylcellulose may be used. Phthalic acid.
  • a dye or pigment can be added to the coating of the tablet or tablet core. For example, a combination for identifying or for characterizing the dosage of an active ingredient.
  • the present invention also provides a method of treating ulcerative colitis comprising administering to a subject in need of treatment a compound of the invention or a pharmaceutical composition comprising a compound of the invention.
  • Methods of administration include, but are not limited to, various methods of administration well known in the art, which can be determined based on the actual circumstances of the patient. These methods include, but are not limited to, parenteral, subcutaneous, intravenous, intramuscular, intraperitoneal, transdermal, buccal, intrathecal, intracranial, nasal or topical routes of administration.
  • the invention also encompasses the use of a compound of the invention in the manufacture of a medicament for the treatment of ulcerative colitis.
  • the compound of the present invention can form a salt or an ester due to the carboxyl group contained therein, thereby forming a prodrug.
  • the present invention has for the first time discovered a series of novel thiazole derivatives having a good therapeutic effect on ulcerative colitis
  • the compound of the present invention is a highly effective and low-toxic therapeutic agent for ulcerative colitis, and thus has important academic value and practical significance.
  • phthaloylbenzoic acid 1.5 g, 10 mmol
  • cesium carbonate 6.5 g, 20 mmol
  • N,N-dimethylformamide DMF
  • O-carboxybenzaldehyde (1 g, 6.7 mmol) was weighed into a 100 mL three-necked flask, and cesium carbonate (4.3 g, 13.3 mmol), potassium iodide (1.1 g, 6.7 mmol), N,N-dimethylformamide (DMF) was added. 15mL, chloromethyl pivalate (1.5g, 10mmol) was added under electromagnetic stirring, and the temperature was raised to 70 ° C by heating, and the reaction was carried out for 20 hours. The reaction was terminated by TLC, the heating was stopped, 20 mL of water was added to the reaction flask, and the reaction solution was used.
  • the synthesis method is the same as that of A1.
  • the reaction is carried out by reverse lysis (the crude product is dissolved completely with ethyl acetate, petroleum ether is added under electromagnetic stirring, rapidly stirred, and the solid is precipitated, and filtered by suction) to obtain a pale yellow powder of 180 mg. The rate is 49%.
  • SPF-class BALB/c male mice each weighing 18-22 g, were purchased from Shanghai Slack Laboratory Animal Co., Ltd., and were used for experiments 7 days after adaptive feeding.
  • TNBS 2,4,6-trinitrobenzenesulfonic acid
  • SASP sulfasalazine tablets
  • anhydrous ethanol was purchased from Shanghai Titan Technology.
  • chloral hydrate was purchased from Jiangsu Qiangsheng Functional Chemical Co., Ltd.
  • sodium chloride was purchased from Sinopharm Chemical Reagent Co., Ltd.
  • 37% ⁇ 40% formaldehyde solution was purchased from Shanghai Lingfeng Chemical Reagent Co., Ltd.
  • carboxymethyl fiber Sodium was purchased from Shanghai Jingchun Biochemical Technology Co., Ltd.
  • paraffin was purchased from Leica, Germany
  • xylene was purchased from Shanghai Lingfeng Chemical Reagent Co., Ltd.
  • hematoxylin and eosin (H&E) staining kit was purchased from Biyuntian Biotechnology Research Institute.
  • Hydrochloric acid was purchased from Shanghai Lingfeng Chemical Reagent Co., Ltd.
  • neutral gum was purchased from Sinopharm Chemical Reagent Co., Ltd.
  • the electronic balance was purchased from Shanghai Jingtian Electronic Instrument Co., Ltd.
  • the precision electronic balance was purchased from METTLER TOLEDO
  • the vernier caliper was purchased from Harbin Measuring and Cutting Tool Group Co., Ltd.
  • the microscope was purchased from Nikon
  • the paraffin slicer was purchased from Leica, Germany.
  • the electric thermostat blast drying oven was purchased from Shanghai Yiheng Scientific Instrument Co., Ltd.
  • mice BALB/c male mice were housed in an animal room with a temperature of 20 to 25 ° C and a relative humidity of 50 to 60%, and white and dark for 12 hours each, and were free to eat and drink.
  • mice were randomly divided into normal control group, trinitrobenzenesulfonic acid (TNBS) model group, drug treatment group and positive drug control group (sulfasalazine 500 mg/kg).
  • TNBS trinitrobenzenesulfonic acid
  • mice were randomly divided into four groups, and the grouping and daily administration were as shown in Table 1:
  • Table 1 Grouping of mice and daily dosing
  • mice in the group were shaved (2 x 2 cm in area) and sensitized with 1% TNBS (dissolved in 50% ethanol). On the 7th day, the mice were fasted to avoid water. After 24 hours, 0.1 ml of 10% chloral hydrate was intraperitoneally injected and 2% TNBS was enema to establish a mouse model of ulcerative colitis. Rat perfused needle was inserted into the colon of mice, the tip of the needle was about 3 to 3.5 cm from the anus, and 0.1 ml of modeling solution was injected (5% TNBS and deionized water were mixed at a volume ratio of 4:1, and the resulting solution was further mixed with absolute ethanol. Mixed by volume ratio 1:1).
  • the normal control group was injected with 0.12 ml of a solvent (50% ethanol solution) in the same manner.
  • Two modeling mice were randomly sacrificed after 24 hours of modeling, and changes in colon were observed to confirm whether the modeling was successful.
  • mice 24 hours after the last administration, the mice were sacrificed by cervical dislocation, the peritoneum was cut open, the whole colon was taken, the intestine was cut longitudinally along the mesenteric margin, rinsed with ice physiological saline, placed on ice, and the pathological changes of the colon were observed and then removed. Fat and mesentery, blot the water with filter paper, weigh the weight of the colon and measure its length. The cells were fixed in 4% formaldehyde for more than 24 hours, and the tissue was dehydrated, embedded in paraffin, and stained with H&E, and the pathological sections were observed and evaluated.
  • the mental state, fur color, and activity of the mice were observed and recorded daily.
  • the body weight of the mice was weighed daily to observe the stool traits.
  • the body weight index according to the disease activity index (DAI) was as follows:
  • Weight constant weight, 0 points; body weight decreased by 1% to 5%, 1 point; decreased by 6% to 10%, 2 points; 11% to 15%, 3 points; more than 15%, 4 points.
  • the weight of the colon was measured with a precision electronic balance, the length was measured with a vernier caliper, and the ratio of colon weight/length was calculated.
  • the pathological changes of colon tissue were observed under light microscope, and the score was recorded.
  • the rating criteria are as follows:
  • Epithelial cells normal morphology, 0 points; loss of goblet cells, 1 point; large loss of goblet cells, 2 points; loss of crypt cells, 3 points; large loss of crypt cells, 4 points.
  • the TNBS model group began to change stool characteristics on the 3rd day after modeling, and there were symptoms of diarrhea. After the 4th day, diarrhea, bloody stools, listlessness, decreased activity, decreased diet, loose and dull fur, weight Significant decline; drug treatment group and positive drug control group, diarrhea symptoms appeared on the third day of modeling, diarrhea, bloody stools, mental dysfunction, decreased activity, decreased diet, dull fur, weight loss, but with TNBS model after the fourth day Compared with the group, the symptoms are lighter. The weight loss is shown in Table 2:
  • the weight/length ratio of the TNBS model group was significantly larger than that of the normal control group, indicating that the hyperemia and swelling of the model group were more serious, while the weight/length ratio of the drug treatment group and the positive drug control group was significantly lower than that of the model group, indicating that the compound of the present invention and the positive drug willow.
  • Azone can reverse the colonic congestion and edema induced by TNBS in mice.
  • the compound of the present invention has excellent therapeutic activity against ulcerative colitis, and some of the compounds have therapeutic activity against ulcerative colitis which is comparable to that of the positive control drug. Therefore, it lays a new material foundation for the development of a new drug for the treatment of ulcerative colitis with low toxicity, high efficiency and safety.

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Abstract

本发明涉及新型噻唑类衍生物在治疗溃疡性结肠炎以及制备治疗溃疡性结肠炎的药物中的应用。具体而言，本发明涉及下式I所示的化合物、含有下式I化合物的药物组合物在治疗溃疡性结肠炎以及制备治疗溃疡性结肠炎的药物中的用途。

Description

新型噻唑类衍生物在治疗炎性肠病中的应用 技术领域
本发明涉及药物化学领域；具体地说，本发明涉及新型的噻唑类衍生物在制备治疗溃疡性结肠炎的药物以及在治疗炎性肠病，特别是溃疡性结肠炎中的应用。
背景技术
溃疡性结肠炎(ulcerative colitis，UC)又称慢性非特异性溃疡性结肠炎，是一种原因不明的肠道粘膜的慢性炎症和溃疡性病变，以累及直肠粘膜、乙状结肠粘膜为主，也可逆行至左半结肠、右半结肠，甚至侵犯到全结肠和末端回肠，与克罗恩病(crohn disease，CD)共称为炎性肠病(inflammatory bowel disease，IBD)。其临床表现主要是腹泻、粘液脓血便、腹痛。病情轻重不等，多呈反复发作的慢性病程。本病可以发生在任何年龄，多见于20-40岁，亦可见于儿童或老年。男女发病率无显著差别。病理特点是粘膜的弥漫性炎症，固有膜内弥漫性淋巴细胞、浆细胞、单核细胞等炎细胞浸润，活动期有大量中性粒细胞和嗜酸性粒细胞浸润，可发生隐窝炎和隐窝脓肿。早在上个世纪，临床医生就对溃疡性结肠炎有所了解，大多数学者认为溃疡性结肠炎是由多因素相互作用所致，主要包括感染、免疫、遗传、环境及精神心理等因素。然而迄今为止，对该病的特异性致病因素和发病机制尚未明确。多年来治疗方法无明显进展，疗效也不令人满意。
目前，针对溃疡性结肠炎的药物并不多，主要有氨基水杨酸、柳氮磺胺吡啶、奥萨拉秦、美沙拉秦，还有免疫抑制剂和激素类药物。尽管这些药物具有一定治疗效果，但也产生了严重的副作用。因此，溃疡性结肠炎被认为是严重影响人们生活质量，且极难治愈的疾病，其特效药物研发受到世界范围内医药公司重视。
因此，本领域急需寻找新型的、更高效、更安全以及成药性更好的溃疡性结肠炎治疗药物。
发明内容
本发明的目的在于提供新型的、更高效、更安全的具备炎性肠病，特别是溃疡性结肠炎抑制活性的化合物，以便用作溃疡性结肠炎的治疗药物。
在第一方面，本发明提供式I所示化合物或其药学上可接受的盐或酯在制备治疗炎性肠病的药物中的用途：
Figure PCTCN2017084749-appb-000001
式中，
R1选自：H、取代或未取代的C1-C6烷基、C3-C6环烷基；独立
R2独立选自：H、卤素、取代或未取代的C1-C6烷基、取代或未取代的C1-C6烷氧基、CN、NO2、羟基、NRaRb；
Ra、Rb可独立选自H或C1-C6烷基
R3选自：H、取代或未取代的C1-C6烷基；
R4选自：H、卤素；
m为0～4的整数；
n为0～5的整数。
在具体的实施方式中，所述炎性肠病是溃疡性结肠炎或克罗恩病；优选溃疡性结肠炎。
在具体的实施方式中，式I中，R1选自：H、取代或未取代的C1-C6烷基；R2选自：H、卤素、取代或未取代的C1-C6烷基、取代或未取代的C1-C6烷氧基；R3选自：H、取代或未取代的C1-C6烷基；R4选自：H、卤素；m为0～2的整数；n为0～2的整数。
在具体的实施方式中，所述化合物如式II所示：
Figure PCTCN2017084749-appb-000002
式中，
R1选自：H、取代或未取代的C1-C3烷基；
R5和R6独立选自：H、卤素、取代或未取代的C1-C3烷基；
R3选自：H、取代或未取代的C1-C3烷基；
R7、R8和R9独立选自：H、卤素。
在具体的实施方式中，式I所示化合物或其药学上可接受的盐或酯是选自下组的 化合物或其药学上可接受的盐或酯：
Figure PCTCN2017084749-appb-000003
Figure PCTCN2017084749-appb-000004
Figure PCTCN2017084749-appb-000005
Figure PCTCN2017084749-appb-000006
在具体的实施方式中，所述化合物是选自下组的化合物：
Figure PCTCN2017084749-appb-000007
在具体的实施方式中，所述化合物是选自下组的化合物：
Figure PCTCN2017084749-appb-000008
Figure PCTCN2017084749-appb-000009
在第二方面，本发明提供一种药物组合物在制备用于治疗炎性肠病的药物中的用途，所述药物组合物含有本发明第一方面所述的化合物或其药学上可接受的盐或酯，以及药学上可接受的载体或赋形剂。
在优选的实施方式中，所述炎性肠病包括但不限于溃疡性结肠炎或克罗恩病；优选溃疡性结肠炎。
在第三方面，本发明提供一种药物组合物，所述药物组合物用于治疗炎性肠病并且含有本发明第一方面所述的化合物或其药学上可接受的盐或酯，以及药学上可接受的载体或赋形剂。
在优选的实施方式中，所述炎性肠病包括但不限于溃疡性结肠炎或克罗恩病；优选溃疡性结肠炎。
在优选的实施方式中，所述药物组合物是适于口服的剂型，包括但不限于片剂、溶液剂、混悬液、胶囊剂、颗粒剂、粉剂。
在第四方面，本发明提供一种治疗炎性肠病的方法，所述方法包括将本发明第一方面所述的化合物或本发明第三方面所述的药物组合物给予需要治疗炎性肠病的对象。
在优选的实施方式中，所述炎性肠病包括但不限于溃疡性结肠炎或克罗恩病；优选溃疡性结肠炎。
应理解，在本发明范围内中，本发明的上述各技术特征和在下文(如实施例)中具体描述的各技术特征之间都可以互相组合，从而构成新的或优选的技术方案。限于篇幅，在此不再一一累述。
附图说明
图1显示了正常对照组、模型组、药物治疗组和阳性对照组中结肠大体形态损伤情况与结肠组织的病理学观察结果(化合物1的应用效果图)。
图2、化合物16(图中312)对TNBS介导的UC模型小鼠的一般情况的影响(对BALB/c小鼠治疗期间每天称量小鼠体重，数据用平均值±SEM的方式表示，#P<0.05,##P<0.01,###P<0.001，与正常对照组相比。*P<0.05,**P<，与模型组相比。)(正常对照组小鼠观察大便无腹泻或便血等异常情况；经TNBS灌肠造模的小鼠大便性状有不同程度的变化：如稀便或血便等。从实验第3天开始，除正常对照组以外的各组BALB/c小鼠陆续出现死亡，其中模型组小鼠腹泻情况和死亡情况最为严重，实验第8天死亡率约为50％。给予柳氮磺胺嘧啶治疗的阳性药对照组小鼠腹泻情况较轻，仅在实验第8天出现一只小鼠死亡。化合物16的三个不同剂量给药组小鼠的腹泻状况跟模型组小鼠相比较有所缓解)
图3、化合物16(图中312)对TNBS介导的UC模型小鼠结肠质量的影响，A图表示不同实验组那个具有代表性结肠照片。B图表示每个实验组中结肠重量与长度的比值。#p<0.05，与对照组相比。*p<0.05，**p<0.01，与模型组相比。从图可知，建立UC模型后，模型组小鼠结肠发生严重病变，给予不同剂量的化合物312(5mg/kg、20mg/kg、50mg/kg)和阳性药可以不同程度地改善结肠溃疡病变程度和内脏黏连的状况，且5mg/kg的312对结肠病变改善效果最佳。
图4、化合物16(图中312)对TNBS诱导的UC模型小鼠结肠组织病理学的影响。结肠组织切片HE染色的代表照片，放大倍数为40。结果显示，正常对照组小鼠结肠组织具有完整的结构，建立UC模型后，模型组小鼠结肠组织完整性被严重破坏，局部出现糜烂。给予阳性药SASP治 疗后，在很大程度上逆转了上述病变；给予化合物16治疗后，跟模型组相比，黏膜完整性已有较大程度的改善，给予5mg/kg和50mg/kg比给予20mg/kg的化合物16效果更理想，其中低剂量5mg/kg与阳性药作用相近。
具体实施方式
发明人经过广泛而深入的研究，出乎意料地发现了一系列具备较好的溃疡性结肠炎治疗效果的噻唑类衍生物，它们的结构骨架完全不同于文献报道过的溃疡性结肠炎治疗药物(例如，氨基水杨酸、柳氮磺胺吡啶、奥萨拉秦、美沙拉秦)。并且这些化合物经实验动物验证显示出显著的溃疡性结肠炎治疗活性。在此基础上完成了本发明。
术语定义
本文中涉及到的一些基团定义如下：
本文中，“烷基”是指碳链长度为1-10个碳原子的饱和的支链或直链烷基，优选的烷基包括长2-8个碳原子、1-6个、1-4个碳原子、1-3个碳原子不等的烷基。烷基的例子包括但不限于甲基、乙基、正丙基、异丙基、正丁基、异丁基、庚基等。烷基可以被1个或多个取代基取代，例如被卤素或卤代烷基取代。例如，烷基可以是被1-4个氟原子取代的烷基，例如三氟甲基，或者烷基可以是被氟代烷基取代的烷基。
本文中，“环烷基”是指含有脂环结构的饱和烷基，例如，C3-C6环烷基。在具体的实施方式中，所述环烷基包括但不限于：环丙基、环丁基、环戊基或环己基。本文所述的环烷基可以是取代或未取代的，包括但不限于被一个或多个卤素原子，例如氟原子取代。
本文中，“氨基”是指结构式为“NRxRy”的基团，其中，Rx和Ry可独立选自H或C1-C3烷基或C1-C3卤代烷基。在具体的实施方式中，本文所述的“氨基”是指NH2。
在本文中，“卤素”是指氟、氯、溴和碘。在优选的实施方式中，卤素是氯或氟；更优选氟。
本发明的化合物
本发明人出乎意料地发现了一系列结构骨架全新的具备较好的炎性肠病，特别是溃疡性结肠炎治疗效果的噻唑类衍生物。这些化合物在动物实验中显示出显著的溃疡性结肠炎治疗活性。
在具体的实施方式中，本发明的化合物是式I所示化合物或其药学上可接受的盐或酯：
Figure PCTCN2017084749-appb-000010
式中，
R1选自：H、取代或未取代的C1-C6烷基、C3-C6环烷基；独立
R2独立选自：H、卤素、取代或未取代的C1-C6烷基、取代或未取代的C1-C6烷氧基、CN、NO2、羟基、NRaRb；
Ra、Rb可独立选自H或C1-C6烷基
R3选自：H、取代或未取代的C1-C6烷基；
R4选自：H、卤素；
m为0～4的整数；
n为0～5的整数。
在优选的实施方式中，R1选自：H、取代或未取代的C1-C6烷基；R2选自：H、卤素(优选F)、取代(优选F取代的)或未取代的C1-C6烷基(优选C1-C3烷基)、取代或未取代的C1-C6烷氧基；R3选自：H、取代或未取代的C1-C6烷基(优选C1-C3烷基)；R4选自：H、卤素(优选Cl)；m为0～2的整数；n为0～2的整数。
在进一步优选的实施方式中，本发明的化合物如式II所示：
Figure PCTCN2017084749-appb-000011
式中，
R1选自：H、取代或未取代的C1-C3烷基；
R5和R6独立选自：H、卤素、取代(优选F取代的)或未取代的C1-C3烷基(优选甲基)；
R3选自：H、取代或未取代的C1-C3烷基；
R7、R8和R9独立选自：H、卤素(优选Cl)。
在上述化合物的基础上，本发明进一步提供一种药物组合物，该组合物含有治疗 有效量的本发明化合物或其药学上可接受的盐或酯，以及药学上可接受的载体或赋形剂。
本发明化合物的药学上可接受的盐的例子包括但不限于无机和有机酸盐，例如盐酸盐、氢溴酸盐、硫酸盐、柠檬酸盐、乳酸盐、酒石酸盐、马来酸盐、富马酸盐、扁桃酸盐和草酸盐；以及与碱例如钠羟基、三(羟基甲基)胺基甲烷(TRIS，胺丁三醇)和N-甲基葡糖胺形成的无机和有机碱盐。
本发明化合物的药学上可接受的酯为式I化合物的羧基被酯化的化合物，包括但不限于：式I化合物的取代或未取代的C1-C10烷基或环烷基的酯、
Figure PCTCN2017084749-appb-000012
基团的酯、
Figure PCTCN2017084749-appb-000013
基团的酯，其中，所述取代指被选自下组的基团取代：苯基、卤素。代表性地，本发明化合物的药学上可接受的酯选自下组：
Figure PCTCN2017084749-appb-000014
Figure PCTCN2017084749-appb-000015
Figure PCTCN2017084749-appb-000016
虽然每个人的需求各不相同，本领域技术人员可确定本发明药物组合物中每种活性成分的最佳剂量。一般情况下，本发明的化合物或其药学上可接受的盐，对哺乳动物每天口服给药，药量按照约0.0025到50毫克/公斤体重。但最好是每公斤口服给药约0.01到10毫克。例如，单位口服剂量可以包括约0.01到50毫克，最好是约0.1到10毫克的本发明化合物。单位剂量可给予一次或多次，每天为一片或多片，每片含有约0.1到50毫克，合宜地约0.25到10毫克的本发明化合物或其溶剂化物。
本发明的药物组合物可被配制成适合各种给药途径的制剂形式，包括但不限于被配制成用于肠外，皮下，静脉，肌肉，腹腔内，透皮，口腔，鞘内，颅内，鼻腔或外用途径给药的形式，用于治疗肿瘤和其他疾病。给药量是有效地改善或消除一个或多个病症的药量。对于特定疾病的治疗，有效量是足以改善或以某些方式减轻与疾病有关的症状的药量。这样的药量可作为单一剂量施用，或者可依据有效的治疗方案给药。给药量也许可治愈疾病，但是给药通常是为了改善疾病的症状。一般需要反复给药来实现所需的症状改善。药的剂量将根据病人的年龄，健康与体重，并行治疗的种类，治疗的频率，以及所需治疗效益来决定。
本发明的药物制剂可以给予任何哺乳动物，只要他们能获得本发明化合物的治疗效果。在这些哺乳动物中最为重要的是人类。本发明的化合物或其药物组合物可用于治疗溃疡性结肠炎。
本发明的药物制剂可用已知的方式制造。例如，由传统的混合，制粒，制锭，溶解，或冷冻干燥过程制造。制造口服制剂时，可结合固体辅料和活性化合物，选择性研磨混合物。如果需要或必要时加入适量助剂后，加工颗粒混合物，获得片剂或锭剂芯。
合适的辅料特别是填料，例如糖类如乳糖或蔗糖，甘露醇或山梨醇；纤维素制剂或钙磷酸盐，例如磷酸三钙或磷酸氢钙；以及粘结剂，例如淀粉糊，包括玉米淀粉，小麦淀粉，大米淀粉，马铃薯淀粉，明胶，黄芪胶，甲基纤维素，羟丙基甲基纤维素，羧甲基纤维素钠，或聚乙烯吡咯烷酮。如果需要，可增加崩解剂，比如上面提到的淀粉，以及羧甲基淀粉，交联聚乙烯吡咯烷酮，琼脂，或褐藻酸或其盐，如海藻酸钠。辅助剂特别是流动调节剂和润滑剂，例如，硅石，滑石，硬脂酸盐类，如镁硬脂酸钙，硬脂酸或聚乙二醇。如果需要，可以給锭剂核芯提供可以抵抗胃液的合适包衣。为此，可以应用浓缩糖类溶液。此类溶液可以含有阿拉伯树胶，滑石，聚乙烯吡咯烷酮，聚乙二醇和/或二氧化钛，漆溶液和合适的有机溶剂或溶剂混合物。为了制备耐胃液的包衣，可使用适当的纤维素溶液，例如醋酸纤维素邻苯二甲酸或羟丙基甲基纤维素邻 苯二甲酸。可向药片或锭剂核芯的包衣加入染料或色素。例如，用于识别或为了表征活性成分剂量的组合。
因此，本发明还提供一种治疗溃疡性结肠炎的方法，该方法包括给予需要治疗的对象以本发明的化合物或包含本发明化合物的药物组合物。
给药方法包括但不限于本领域周知的各种给药方法，可根据患者的实际情况加以确定。这些方法包括但不限于肠外、皮下、静脉、肌肉、腹腔内、透皮、口腔、鞘内、颅内、鼻腔或外用途径给药。
本发明也包括本发明化合物在制备治疗溃疡性结肠炎的药物中的用途。
此外，本领域技术人员基于本领域的公知常识和本发明的内容可以知晓，本发明化合物因其中所含的羧基而能形成盐或酯，进而可以形成前药。
本发明的优点
1.本发明首次发现了一系列结构全新的具备较好的溃疡性结肠炎治疗效果的噻唑类衍生物；
2.本发明的化合物是高效、低毒的溃疡性结肠炎治疗剂，从而具备很重要的学术价值与现实意义。
以下结合具体实施案例对本发明的技术方案进一步描述，但以下实施例不构成对本发明的限制，所有依据本发明的原理和技术手段采用的各种施用方法，均属于本发明范围。下列实施例中未注明具体条件的实验方法，通常按照常规条件，或按照制造厂商所建议的条件。除非另外说明，否则百分比和份数按重量计算。
实施例1：
(E)-4-(2-氯苯基)-2-[1-甲基-2-(2-羧基苄叉基)肼基]噻唑1
Figure PCTCN2017084749-appb-000017
2-甲基氨基硫脲(1-1)
称取2.5g(17.3mmol)甲基肼硫酸盐于250ml单口瓶中，加入100ml乙醇，搅拌下加入1.6g(20.8mmol)硫氰酸铵，加热至回流，反应72h后，将反应液冷至室温，抽滤，滤液旋干硅胶柱层析(DCM/MeOH＝40:1)，分第二个副产物，得白色粉末状固体0.63g，产率34.2％。1H NMR(400MHz,DMSO-d6,ppm)δ7.36(s,2H),4.89(s,2H),3.41(s,3H).GC-MS(EI)calcd for C2H7N3S[M]+105.0,found 105.0.
2-甲基-1-(2-羧基苄差基)氨基硫脲(1-2)
称取80mg(0.76mmol)化合物(1)于50ml单口瓶中，加入20ml乙醇，搅拌下加入邻羧基苯甲醛114mg(0.76mmol)，加热至回流，TLC监测反应至原料转化完全，将反应液冷至室温，旋干溶剂，硅胶柱层析(DCM/MeOH＝120:1)，得白色粉末状固体100mg，产率56％。1H NMR(400MHz,DMSO-d6,ppm)δ13.36(br,1H),8.56(s,1H),8.51(s,1H),8.31(d,J＝7.8Hz,1H),8.25(s,1H),7.88(d,J＝8Hz,1H),7.59(t,J＝7.2Hz,1H),7.50(t,J＝7.2Hz,1H),3.77(s,3H).LC-MS(ESI)calcd for C10H12N3O2S[M+H]+238.1,found 238.1.
(E)-4-(2-氯苯基)-2-[1-甲基-2-(2-羧基苄叉基)肼基]噻唑1
称取100mg(0.42mmol)化合物(2)于50ml单口瓶中，加入10ml乙醇，搅拌下加入65μL(0.42mmol)2’-氯-2-溴苯乙酮，升温至回流，TLC跟踪反应至原料转化完全，将反应液冷至室温，旋干溶剂，硅胶柱层析(DCM/MeOH＝120:1)，得黄色粉末状固体106mg，产率67.9％，Mp.210.4-212.0℃。1H NMR(400MHz,DMSO-d6,ppm)δ13.30(s,1H),8.62(s,1H),8.02(d,J＝7.6Hz,1H),7.98–7.90(m,2H),7.66(t,J＝7.6Hz,1H),7.56–7.49(m,2H),7.47(s,1H),7.43(td,J1＝7.4Hz,J2＝1.2Hz,1H),7.36(td,J1＝7.6Hz,J2＝1.6Hz,1H),3.68(s,3H).13C NMR(125MHz,DMSO-d6,ppm)δ168.98,168.57,147.11,136.94,135.17,133.42,132.48,131.51,131.09,130.98,130.75, 130.16,129.45,129.19,127.63,126.51,111.41,32.88.HRMS(ESI)calcd for C18H15N3O2SCl[M+H]+372.0574,found 372.0575.
采取类似的方法，本发明人利用相应的起始材料进一步合成了以下化合物：
(E)-4-(2-氯苯基)-2-[1-乙基-2-(2-羧基苄叉基)肼基]噻唑2
Figure PCTCN2017084749-appb-000018
Mp.203.3-204.1℃.1H NMR(500MHz,DMSO-d6,ppm)δ13.32(br,1H),8.64(s,1H),7.98–7.91(m,3H),7.66(t,J＝7.5Hz,1H),7.56–7.46(m,2H),7.45–7.39(m,2H),7.34(t,J＝6.8Hz,1H),4.32(q,J＝6.8Hz,2H),1.27(t,J＝7.0Hz,3H).13C NMR(125MHz,DMSO-d6,ppm)δ168.56,168.36,147.34,137.00,135.42,133.53,132.50,131.53,131.12,130.92,130.72,130.22,129.42,129.19,127.62,126.62,111.22,40.38,10.33.HRMS(ESI)calcd for C19H17N3O2SCl[M+H]+386.0730,found 386.0728.
(E)-4-(2-氯苯基)-2-[1-丙基-2-(2-羧基苄叉基)肼基]噻唑3
Figure PCTCN2017084749-appb-000019
Mp.172.6-173.4℃.1H NMR(400MHz,DMSO-d6,ppm)δ13.35(br,1H),8.65(s,1H),7.98(d,J＝8.0Hz,1H),7.95-7.92(m,2H),7.67(t,J＝7.2Hz,1H),7.54(d,J＝8.0Hz,1H),7.5(t,J＝7.6Hz,1H),7.45-7.42(m,2H),7.36(t,J1＝7.6Hz,J2＝1.6Hz,1H),4.25(t,J＝7.2Hz,2H),1.82-1.73(m,2H),0.97(t,J＝7.4Hz,1H).13C NMR(100MHz,DMSO-d6,ppm)δ168.54,168.22 146.95,136.63,134.96,133.17,132.11,131.10,130.74,130.55,130.34,129.82,129.04,128.80,127.26,126.09,110.77,46.31,17.98,11.17.HRMS(ESI)calcd for C20H19N3O2SCl[M+H]+400.0887,found 400.0879.
(E)-4-(2-氯苯基)-2-[1-异丙基-2-(2-羧基苄叉基)肼基]噻唑4
Figure PCTCN2017084749-appb-000020
Mp.185.4-187.0℃.1H NMR(400MHz,DMSO-d6,ppm)δ13.35(br,1H),8.91(s,1H),7.99(d,J＝8.0Hz,1H),7.95-7.92(m,2H),7.67(t,J＝7.6Hz,1H),7.54(d,J＝8.0Hz,1H),7.52(t,J＝7.6Hz,1H),7.46-7.42(m,2H),7.36(td,J1＝7.6Hz,J2＝1.6Hz,1H),5.22-5.11(m,1H),1.57(d,J＝6.8Hz,6H).13C NMR(100MHz,DMSO-d6,ppm)δ168.22,168.21,146.90,137.32,135.23,133.14,132.07,131.06,130.64,130.58,130.39,129.88,128.97,128.78,127.28,125.91,111.12,49.61,18.07,18.07.HRMS(ESI)calcdfor C20H19N3O2SCl[M+H]+400.0887,found 400.0885.
(E)-4-(2-氯苯基)-2-[1-(2-丁基)-2-(2-羧基苄叉基)肼基]噻唑5
Figure PCTCN2017084749-appb-000021
Mp.170.5-170.6℃.1H NMR(400MHz,DMSO-d6,ppm)δ13.32(br,1H),8.91(s,1H),7.99(d,J＝8.0Hz,1H),7.95-7.90(m,2H),7.67(t,J1＝7.6Hz,1H),7.55-7.48(m,2H),7.46-7.42(m,2H),7.36(td,J1＝7.6Hz,J2＝1.6Hz,1H),5.00-4.93(m,1H),2.35-2.24(m,1H),1.92-1.81(m,1H),1.54(d,J＝6.8Hz,3H),0.88(t,J＝7.2Hz,3H).13C NMR(100MHz,DMSO-d6,ppm)δ168.66,168.19,146.97,137.03,135.24,133.19,132.13,131.04,130.67,130.58,130.39,129.70,128.98,128.77,127.29,125.91,111.04,55.67,25.26,16.33,11.14.HRMS(ESI)calcd for C21H21N3O2SCl[M+H]+414.1043,found 414.1029.
(E)-4-(2-氯苯基)-2-[1-(2-戊基)-2-(2-羧基苄叉基)肼基]噻唑6
Figure PCTCN2017084749-appb-000022
Mp.146.7-147.0℃.1H NMR(400MHz,DMSO-d6,ppm)δ13.35(br,1H),8.91(s,1H),7.98(d,J＝7.6Hz,1H),7.94(d,J＝8.0Hz,1H),7.91(dd,J1＝7.6Hz,J2＝1.6Hz,1H),7.67(t,J＝7.6Hz,1H),7.53(d,J＝8.0Hz,1H),7.49(t,J＝7.6Hz,1H),7.46-7.42(m,2H),7.36(td,J1＝7.6Hz,J2＝1.6Hz,1H),5.14-5.06(m,1H),2.34-2.25(m,1H),1.82-1.73(m,1H),1.53(d,J＝6.8Hz,3H),1.34-1.24(m,2H),0.89(t,J＝7.4Hz,1H).13C NMR(100MHz,DMSO-d6,ppm)δ168.68,168.20,146.98,137.04,135.30,133.20,132.14,131.00,130.68,130.59,130.40,129.68,128.99,127.30,125.90,111.09,53.81,34.20,19.46,16.46,13.60.HRMS(ESI)calcd for C22H23N3O2SCl[M+H]+428.1200,found 428.1193.
(E)-4-(2-氯苯基)-2-[1-羟乙基-2-(2-羧基苄叉基)肼基]噻唑7
Figure PCTCN2017084749-appb-000023
Mp.187.9-188.9℃.1H NMR(400MHz,DMSO-d6,ppm)δ13.24(br,1H),8.75(s,1H),7.98-7.01(m,3H),7.66(t,J＝7.6Hz,1H),7.54(d,J＝8.0Hz,1H),7.49(t,J＝7.6Hz,1H),7.45-7.41(m,2H),7.36(td,J1＝7.6Hz,J2＝1.6Hz,1H),4.35(t,J＝6.4Hz,2H),3.76(t,J＝6.4Hz,2H).13C NMR(100MHz,DMSO-d6,ppm)δ168.64,168.23,146.87,136.89,134.91,133.15,131.99,131.19,130.71,130.46,130.32,130.16,129.04,128.79,127.22,126.28,110.78,56.24,47.46.HRMS(ESI)calcd for C19H17N3O3SCl[M+H]+ 402.0679,found 402.0678.
(E)-4-(2-氯苯基)-2-[1-甲基-2-(4-三氟甲基-2-羧基苄叉基)肼基]噻唑8
Figure PCTCN2017084749-appb-000024
Mp.209.3-210.7℃.1H NMR(400MHz,DMSO-d6,ppm)δ14.02(br,1H),8.66(s,1H),8.21(d,J＝8.0Hz,1H),8.17(s,1H),8.02(d,J＝8.0Hz,1H),7.95(d,J＝7.6Hz,1H),7.56-7.53(m,2H),7.43(t,J＝7.6Hz,1H),7.37(t,J＝7.6Hz,1H),3.70(s,3H).13C NMR(100MHz,DMSO-d6,ppm)δ168.33,167.17,146.79,138.38,135.01,132.91,131.12,130.70,130.37,129.14,128.57,128.25,128.20,127.26,127.04,125.10,122.40,111.51,32.68.HRMS(ESI)calcd for C19H14N3O2SClF3[M+H]+440.0447,found440.0433.
(E)-4-(2-氯苯基)-2-[1-甲基-2-(4-甲基-2-羧基苄叉基)肼基]噻唑9
Figure PCTCN2017084749-appb-000025
Mp.231.3-232.5℃.1H NMR(400MHz,DMSO-d6,ppm)δ13.30(br,1H),8.59(s,1H),7.96(d,J＝7.6Hz,1H),7.92(d,J＝8.0Hz,1H),7.75(s,1H),7.54(d,J＝7.6Hz,1H),7.49-7.46(m,2H),7.43(t,J＝7.6Hz,1H),7.36(t,J＝7.6Hz,1H),3.66(s,3H),2.38(s,3H).13C NMR(100MHz,DMSO-d6,ppm)δ168.60,168.35,146.68,138.54,136.64,133.03,132.70,132.07,131.11,130.86,130.67,130.34,129.83,129.00,127.21,126.04,110.84,32.39,20.69.HRMS(ESI)calcd for C19H17N3O2SCl[M+H]+386.0730,found386.0738.
(E)-4-(2-氯苯基)-2-[1-甲基-2-(2-羧基-4-氟苄叉基)肼基]噻唑10
Figure PCTCN2017084749-appb-000026
Mp.228.6-229.7℃.1H NMR(400MHz,DMSO-d6,ppm)δ13.28(br,1H),8.14(s,1H),7.93(dd,J1＝7.6Hz,J2＝1.6Hz,1H),7.61(dd,J1＝7.0Hz,J2＝1.8Hz,1H),7.55-7.49(m,3H),7.44-7.41(m,1H),7.36(td,J1＝7.6Hz,J2＝1.6Hz,1H),3.68(s,3H).13C NMR(100MHz,DMSO-d6,ppm)δ168.55,168.01,159.74(d,1J＝249Hz),146.67,133.68,133.10,131.50,131.12,130.72,130.32,130.10(d,3J＝8.8Hz),129.06,127.23,125.53(d,4J＝3.1Hz),121.75(d,2J＝11.8Hz),118.76(d,2J＝22Hz),111.13,32.28.HRMS(ESI)calcd for C18H14N3O2FSCl[M+H]+390.0479,found 390.0475.
(E)-5-甲基-4-苯基-2-[1-甲基-2-(4-甲基-2-羧基苄叉基)肼基]噻唑11
Figure PCTCN2017084749-appb-000027
Mp.243.2-245.2℃.1H NMR(400MHz,DMSO-d6,ppm)δ13.24(br,1H),8.54(s,1H),7.91(d,J＝8.0Hz,1H),7.73(s,1H),7.64-7.53(m,2H),7.46-7.42(m,3H),7.33(t,J＝7.4Hz,1H),3.60(s,3H),2.44(s,3H),2.38(s,3H).13C NMR(100MHz,DMSO-d6,ppm)δ168.35,165.40,145.43,138.33,135.76,135.16,132.68,132.22,130.86,129.62,128.24,128.24,127.85,127.85,127.06,125.89,119.27,31.85,20.68,12.24.HRMS(ESI)calcd for C20H20N3O2S[M+H]+366.1276,found 366.1273.
(E)-5-甲基-4-苯基-2-[1-甲基-2-(2-羧基苄叉基)肼基]噻唑12
Figure PCTCN2017084749-appb-000028
Mp.222.7-224.9℃.1H NMR(400MHz,DMSO-d6,ppm)δ13.32(br,1H),8.58(s,1H),8.01(d,J＝8.0Hz,1H),7.92(d,J＝7.6Hz,1H),7.68-7.63(m,3H),7.50-7.43(m,3H)，7.34(t,J＝7.4Hz,1H),3.62(s,3H),2.44(s,3H).13C NMR(100MHz,DMSO-d6,ppm)δ168.24,165.36,145.47,135.66,135.13,134.90,131.98,130.58,129.70,128.59,128.25,128.25,127.86,127.86,127.09,125.94,119.44,31.93,12.24.HRMS(ESI)calcd for C19H16N3O2S[M-H]-350.0963,found 350.0951.
(E)-4-(2-氯苯基)-2-[1-甲基-2-(2-羧基苄叉基)肼基]噻唑13
Figure PCTCN2017084749-appb-000029
Mp.208.8-209.8℃.1H NMR(400MHz,DMSO-d6,ppm)δ13.46(br,1H),8.60(s,1H),8.01(d,J＝8.0Hz,1H),7.92(d,J＝8.0Hz,1H),7.64(t,J＝7.4Hz,1H),7.56(d,J＝7.6Hz,1H),7.49-7.39(m,4H),3.56(s,3H),2.15(s,3H).13C NMR(100MHz,DMSO-d6,ppm)δ168.98,166.46,144.56,136.46,135.29,134.54,133.25,132.50,132.31,131.05,130.72,130.26,130.01,129.09,127.49,126.42,121.85,32.46,12.16.HRMS(ESI)calcd for C19H17N3O2SCl[M+H]+386.0730,found 386.0727.
(E)-5-乙基-4-(2-氯苯基)-2-(2-羧基苄叉肼基)噻唑14
Figure PCTCN2017084749-appb-000030
Mp.223.9-223.9℃.1H NMR(500MHz,DMSO-d6,ppm)δ12.57(br,2H),8.79(s,1H),8.01(d,J＝8.0Hz,1H),7.88(d,J＝8.0Hz,1H),7.62(t,J＝7.5Hz,1H),7.56–7.51(m,1H),7.46(t,J＝7.5Hz,1H),7.44-7.36(m,3H),2.49(q,J＝7.5Hz,1H),1.13(t,J＝ 7.5Hz,3H).13C NMR(125MHz,DMSO-d6,ppm)δ169.28,165.92,144.14,140.84,135.89,135.46,133.99,133.03,133.02,131.46,130.81,130.73,130.60,129.73,128.09,127.73,127.01,20.99,17.25.HRMS(ESI)calcd for C19H17N3O2SCl[M+H]+386.0730,found 386.0729.
(E)-5-甲基-4-(2-氯苯基)-2-(2-羧基苄叉肼基)噻唑15
Figure PCTCN2017084749-appb-000031
Mp.217.6-217.8℃.1H NMR(400MHz,DMSO-d6,ppm)δ12.7(br,2H),8.77(s,1H),7.99(d,J＝7.6Hz,1H),7.87(d,J＝7.2Hz,1H),7.62(t,J＝7.4Hz,1H),7.562-7.543(m,1H),7.481-7.397(m,4H),2.14(s,3H).13C NMR(125MHz,DMSO-d6,ppm)δ168.65,165.18,144.49,140.16,135.25,134.56,133.21,132.46,130.83,130.14,130.03,129.11,127.45,126.37,119.49,12.15.HRMS(ESI)calcd for C18H15N3O2SCl[M+H]+372.0574,found 372.0569.
(E)-4-(2-氯苯基)-2-(2-羧基苄叉肼基)噻唑16
Figure PCTCN2017084749-appb-000032
Mp.200.2-200.9℃.1H NMR(400MHz,DMSO-d6,ppm)δ13.2(br,1H),12.4(br,1H),8.82(s,1H),8.00(d,J＝7.6Hz,1H),7.89-7.86(m,2H),7.64(t,J＝7.4Hz,1H),7.53(dd,J1＝7.6Hz,J2＝1.2Hz,1H),7.49(td,J1＝7.6Hz,J2＝1.2Hz,1H),7.42(td,J1＝7.6Hz,J2＝1.2Hz,1H),7.36(m,2H).13C NMR(100MHz,DMSO-d6,ppm)δ168.63,167.77,147.63,140.76,135.08,133.71,132.43,131.54,131.23,130.83,130.83,130.26,129.47,129.28,127.69,126.47,109.26.HRMS(ESI)calcd for C17H13N3O2SCl[M+H]+358.0417,found 358.0417.
(E)-5-甲基-4-苯基-2-(2-羧基苄叉肼基)噻唑17
Figure PCTCN2017084749-appb-000033
Mp.215.9-216.0℃.1H NMR(400MHz,DMSO-d6,ppm)δ12.7(br,2H),8.77(s,1H),7.99(d,J＝7.6Hz,1H),7.87(dd,J1＝8.0Hz,J2＝1.2Hz,1H),7.63-7.59(m,3H),7.48-7.43(m,3H),7.33(t,J＝7.2Hz,1H),2.43(s,3H).13C NMR(125MHz,DMSO-d6,ppm)δ168.66,164.72,146.00,140.05,135.64,135.24,132.36,130.83,130.12,129.08,128.73,128.73,128.35,128.35,127.51,126.33,117.61,12.74.HRMS(ESI)calcd forC18H16N3O2S[M+H]+338.0963,found 338.0954.
(E)-4-苯基-2-(2-羧基苄叉肼基)噻唑18
Figure PCTCN2017084749-appb-000034
Mp.163.8-165.6℃.1H NMR(400MHz,DMSO-d6,ppm)δ13.45(br,1H),12.49(br,1H),8.81(s,1H),7.99(d,J＝7.6Hz,1H),7.87(m,3H),7.63(t,J＝7.4Hz,1H),7.47(t,J＝7.2Hz,1H),7.41(t,J＝7.6Hz,2H),7.33(s,1H),7.30(t,J＝7.4Hz,1H).13C NMR(100MHz,DMSO-d6,ppm)δ168.53,168.53,150.97,140.58,135.02,134.99,132.31,130.74,130.15,129.15,128.96,128.96,127.90,126.34,125.90,125.90,104.20.HRMS(ESI)calcd for C18H16N3O2S[M-H]-322.0650,found 322.0656.
(E)-4-(2,5-二氯苯基)-2-[1-甲基-2-(2-羧基苄叉基)肼基]噻唑19
Figure PCTCN2017084749-appb-000035
Mp.268.4-269.5℃.1H NMR(400MHz,DMSO-d6,ppm)δ13.31(br,1H),8.63(s,1H),8.02-8.00(m,2H),7.93(d,J＝7.2Hz,1H),7.67(t,J＝7.6Hz,1H),7.63(s,1H),7.58(d,J＝7.6Hz,1H),7.50(t,J＝7.2Hz,1H),7.43(dd,J1＝7.2Hz,J2＝2.8Hz,1H),3.68(s,3H).13C NMR(100MHz,DMSO-d6,ppm)δ168.68,168.15,145.13,136.79,134.70,134.21,132.14,132.06,131.85,130.59,130.17,129.76,129.20,128.84,128.58,126.13,112.31,32.49.HRMS(ESI)calcd for C18H14N3O2SCl2[M+H]+406.0184,found 406.0187.
(E)-4-苯基-2-[1-甲基-2-(2-羧基苄叉基)肼基]噻唑20
Figure PCTCN2017084749-appb-000036
Mp.205.6-206.8℃.1H NMR(400MHz,DMSO-d6,ppm)δ13.31(br,1H),8.62(s,1H),8.02(d,J＝7.6Hz,1H),7.94-7.91(m,3H),7.66(t,J＝7.2Hz,1H),7.49(t,J＝7.2Hz,1H),7.45-7.40(m,3H),7.31(t,J＝7.2Hz,1H),3.71(s,3H).13C NMR(100MHz,DMSO-d6,ppm)δ169.45,168.20,150.18,136.35,134.80,134.49,132.07,130.60,129.71,128.76,128.56,128.56,127.59,126.09,125.54,125.54,105.97,32.51.HRMS(ESI)calcd for C18H16N3O2S[M+H]+338.0963,found 338.0963.
(E)-4-(3-氯苯基)-2-[1-甲基-2-(2-羧基苄叉基)肼基]噻唑21
Figure PCTCN2017084749-appb-000037
Mp.244.7-245.4℃.1H NMR(400MHz,DMSO-d6,ppm)δ13.32(br,1H),8.63(s,1H),8.01(d,J＝7.6Hz,1H),7.97(s,1H),7.93(d,J＝7.6Hz,1H),7.88(d,J＝8.0Hz,1H),7.66(t,J＝7.6Hz,1H),7.61(s,1H),7.50(t,J＝7.6Hz,1H),7.45(t,J＝7.6Hz,1H), 7.37(d,J＝8.0Hz,1H),3.71(s,3H).13C NMR(100MHz,DMSO-d6,ppm)δ169.57,168.18,148.56,136.64,136.50,134.72,133.48,132.06,130.59,130.44,129.76,128.81,127.28,126.12,125.16,124.05,107.56,32.52.HRMS(ESI)calcd for C18H15N3O2SCl[M+H]+372.0574,found 372.0574.
(E)-5-甲基-4-(2-氯苯基)-2-[1-(2-戊基)-2-(2-羧基苄叉基)肼基]噻唑22
Figure PCTCN2017084749-appb-000038
Mp.89.9-90.1℃.1H NMR(400MHz,DMSO-d6,ppm)δ13.29(br,1H),8.85(s,1H),7.98(d,J＝8.0Hz,1H),7.92(d,J＝7.6Hz,1H),7.65(t,J＝7.6Hz,1H),7.56-7.54(m,1H),7.51-7.41(m,4H),5.05-4.96(m,1H),2.29-2.22(m,1H),2.16(s,3H),1.75-1.67(m,1H),1.46(d,J＝7.2Hz,3H),1.28-1.23(m,2H),0.87(t,J＝7.6Hz,3H).13C NMR(100MHz,DMSO-d6,ppm)δ168.23,166.06,144.08,136.14,135.46,134.09,132.79,132.09,131.93,130.58,129.64,129.60,129.48,128.56,126.95,125.73,121.62,53.39,34.01,19.41,16.35,13.58,11.52.HRMS(ESI)calcd for C23H25N3O2SCl[M+H]+442.1356,found 442.1354.
(E)-2-((2-(4-(3-甲氧基苯基)噻唑-2-基)亚肼基)甲基)苯甲酸(52)
Figure PCTCN2017084749-appb-000039
熔点：167.3-167.9℃.1H NMR(400MHz,DMSO-d6):δ12.80(s,1H),8.82(s,1H),8.00(d,J＝8.0Hz,1H),7.88(d,J＝7.6Hz,1H),7.62(t,J＝7.6Hz,1H),7.49-7.43(m,3H),7.37(s,1H),7.32(t,J＝8.0Hz,1H),6.88(dd,J1＝8.4Hz，J2＝2.0Hz，1H),3.80(s,3H).13C NMR(100MHz,DMSO-d6):168.67,168.52,159.99,150.90,140.75,136.52,135.04,132.36,130.81,130.40,130.12,129.26,126.43,118.41,113.80,111.31,104.71,55.52.HRMS(ESI)calcd for C18H14N3O3S[M-H]-352.0756,found 352.0754.Purity:95.56％(tR 7.94min).
(E)-2-((2-(4-(3-氨基甲酰基苯基)噻唑-2-基)亚肼基)甲基)苯甲酸(53)
Figure PCTCN2017084749-appb-000040
熔点：297.9-298.4℃.1H NMR(400MHz,DMSO-d6):δ12.20(s,1H),8.83(s,1H),8.39(s,1H),8.05(s,1H),8.00(dd,J1＝7.2Hz，J2＝3.2Hz,2H),7.88(d,J＝7.6Hz,1H),7.80(d,J＝7.6Hz,1H),7.63(t,J＝7.6Hz,1H),7.51-7.46(m,2H),7.41(s,2H).13C NMR(100MHz,DMSO-d6):168.77,168.67,168.44,150.57,140.83,135.19,135.14,135.02,132.40,130.83,130.36,129.31,129.03,128.62,126.91,126.46,125.36,105.00. HRMS(ESI)calcd for C18H15N4O3S[M+H]+367.0865,found 367.0863.Purity:97.89％(tR 7.832min).
(E)-2-((2-(4-(对甲苯基)噻唑-2-基)亚肼基)甲基)苯甲酸(54)
Figure PCTCN2017084749-appb-000041
熔点：225.5-226.1℃.1H NMR(400MHz,DMSO-d6):δ12.80(s,1H),8.83(s,1H),8.00(d,J＝7.2Hz,1H),7.88(d,J＝7.6Hz,1H),7.75(d,J＝8.0Hz,2H),7.62(t,J＝7.6Hz,1H),7.47(t,J＝7.6Hz,1H),7.25(s,1H),7.21(t,J＝8.0Hz,2H),2.32(s,3H).13C NMR(100MHz,DMSO-d6):168.73,168.55,151.12,140.69,137.26,135.04,132.50,132.29,130.80,130.56,129.63,129.22,126.39,125.96,103.37,21.27.HRMS(ESI)calcd for C18H14N3O2S[M-H]-336.0807,found 336.0808.Purity:96.31％(tR 13.17min).
(E)-2-((2-(4-(2,5-二氯苯基)噻唑-2-基)亚肼基)甲基)苯甲酸(55)
Figure PCTCN2017084749-appb-000042
熔点：205.0-206.0℃.1H NMR(400MHz,DMSO-d6):δ12.20(s,1H),8.84(s,1H),7.99(d,J＝7.2Hz,1H),7.95(d,J＝6.4Hz,1H),7.88(d,J＝8.0Hz,1H),7.62(t,J＝7.2Hz,1H),7.57(d,J＝8.4Hz,1H),7.52(s,1H),7.47(t,J＝7.6Hz,1H),7.41(dd,J1＝8.8Hz，J2＝2.8Hz，1H).13C NMR(100MHz,DMSO-d6):168.67,167.90,146.07,141.15,135.02,134.92,132.65,132.33,132.28,130.82,130.74,130.62,129.74,129.35,129.03,126.46,110.57.HRMS(ESI)calcd for C17H10N3O2SCl2[M-H]-389.9871,found 389.9871.Purity:97.66％(tR 9.88min).
(E)-2-((2-(4-(2,5-二氯苯基)噻唑-2-基)亚肼基)甲基)苯甲酸(56)
Figure PCTCN2017084749-appb-000043
熔点：238.1-239.4℃.1H NMR(400MHz,DMSO-d6):δ12.80(s,1H),8.87(s,1H),8.40(s,1H),8.04-8.01(m,2H),7.96-7.89(m,4H),7.64(t,J＝7.6Hz,1H),7.54-7.46(m,4H).13C NMR(100MHz,DMSO-d6):168.75,168.68,151.02,140.83,135.07,133.65,132.92,132.58,132.40,130.85,130.39,129.29,128.63,128.57,128.05,126.90,126.47,124.57,124.43,105.15.HRMS(ESI)calcd for C21H16N3O2S[M+H]+374.0963,found 374.0959.Purity:96.39％(tR 19.11min).
(E)-2-((2-(4-(萘-2-基)噻唑-2-基)亚肼基)甲基)苯甲酸(57)
Figure PCTCN2017084749-appb-000044
熔点：227.1-228.1℃.1H NMR(400MHz,DMSO-d6):δ12.35(s,1H),8.83(s,1H), 8.01(d,J＝6.0Hz,1H),7.96(d,J＝6.8Hz,2H),7.88(d,J＝6.0Hz,1H),7.74-7.71(m,4H),7.64(t,J＝6.4Hz,1H),7.48(t,J＝6.0Hz,3H),7.42(s,1H),7.38(t,J＝6.0Hz,1H).13C NMR(100MHz,DMSO-d6):169.31,169.31,151.40,141.42,140.82,140.22,135.75,134.93,133.11,131.52,130.98,130.12,129.97,128.62,127.99,127.63,127.26,127.14,105.26.HRMS(ESI)calcd for C23H18N3O2S[M+H]+400.1120,found 400.1114.Purity:97.60％(tR 9.16min).
(E)-2-((2-(4-(2-氯苯基)噻唑-2-基)亚肼基)甲基)-6-氟苯甲酸(58)
Figure PCTCN2017084749-appb-000045
熔点：211.2-211.7℃.1H NMR(400MHz,DMSO-d6):δ12.40(s,1H),8.18(s,1H),7.86(dd,J1＝7.6Hz，J2＝1.6Hz，1H),7.69(d,J＝8.0Hz,1H),7.55-7.50(m,2H),7.41(t,J＝7.6Hz,1H),7.37(s,1H),7.35(t,J＝7.6Hz,1H),7.29(t,J＝8.8Hz,1H).13C NMR(100MHz,DMSO-d6):167.56,166.14,160.51,158.06,147.64,138.21,138.17,133.94,133.89,133.68,131.54,131.25,130.83,129.53,127.72,122.04,116.51,116.29,109.58.HRMS(ESI)calcd for C17H10N3O2SClF[M-H]-374.0166,found 374.0164.Purity:97.09％(tR 12.05min).
(E)-2-((2-(4-(2-氯苯基)噻唑-2-基)亚肼基)甲基)-3-氨基苯甲酸(59)
Figure PCTCN2017084749-appb-000046
mp 220.9-221.6℃.1H NMR(400MHz,DMSO-d6):δ12.8(s,1H),11.1(s,1H),7.93(s,1H),7.89(d,J＝7.2Hz,1H),7.62(d,J＝7.6Hz,1H),7.52-7.43(m,3H),7.12(d,J＝7.2Hz,1H),7.08(d,J＝8.4Hz,1H),6.77(s,2H).13C NMR(100MHz,DMSO-d6):δ165.84,155.22,154.28,147.07,146.30,133.99,132.71,131.65,131.51,131.09,130.33,129.73,128.14,121.28,114.49,113.14,111.55.Purity:97.06％(tR 19.58min).
2-甲酰基苯甲酸乙酯1a
Figure PCTCN2017084749-appb-000047
称取邻甲酰基苯甲酸(1.5g,10mmol)于100mL三口瓶中，加入碳酸铯(6.5g,20mmol)，N,N-二甲基甲酰胺(DMF)20mL，电磁搅拌下加入碘乙烷(4.68g,30mmol)，加热升温至70℃，反应4h，TLC监测至反应结束，停止加热，向反应瓶中加入20mL水，反应液用二氯甲烷和水萃取三次，合并有机相，加入无水硫 酸钠干燥，抽滤，滤液加入等质量的硅胶旋干，过柱纯化，得到无色透明液体1.4g，产率79％。1H NMR(400MHz,DMSO-d6,ppm):δ10.41(s,1H),7.92-7.90(m,1H),7.88-7.85(m,1H),7.81-7.77(m,2H),4.40(q,J＝7.2Hz,2H),1.34(t,J＝7.2Hz,3H).LC-MS(ESI)calcd for C10H10O3[M+H]+179.06,found 179.10.
(E)-4-(2-氯苯基)-2-(2-乙氧基甲酰基苄叉肼基)噻唑A1
Figure PCTCN2017084749-appb-000048
称取化合物1a(178mg,1mmol)于50mL两口瓶中，另取化合物3(225mg,1mmol)加入反应瓶中，加入无水乙醇15mL，氮气保护下，电磁搅拌反应4h，TLC检测原料反应完全，抽滤反应液，滤饼用乙醇淋洗，干燥得到近纯产物280mg，过柱纯化后得到淡黄色固体粉末220mg，产率57％。熔点：172.1-172.8℃.1H NMR(400MHz,DMSO-d6,ppm):δ13.30(br,1H),12.05(br,1H),8.84(s,1H),8.02(d,J＝7.6Hz,1H),7.91(t,J＝8.0Hz,2H),7.66(t,J＝7.2Hz,1H),7.55(d,J＝7.6Hz,1H),7.51(t,J＝8Hz,1H),7.44(t,J＝7.2Hz,1H),7.38-7.34(m,2H).13C NMR(100MHz,DMSO-d6,ppm):δ167.92,16.70,147.26,141.03,134.55,133.23,131.94,131.82,131.06,130.70,130.34,128.80,128.74,128.60,127.22,125.96,109.26,61.23,14010.HRMS(ESI)calcd for C19H17N3O2SCl[M+H]+386.0730,found 386.0731.HPLC purity:98.88％,Retention time＝16.14min.
2-甲酰基苯甲酸丙酯2a
Figure PCTCN2017084749-appb-000049
合成方法同1a，得到透明液体1.5g，产率78％。1H NMR(400MHz,DMSO-d6,ppm):δ10.40(s,1H),7.93-7.90(m,1H),7.88-7.84(m,1H),7.81-7.77(m,2H),4.29(t,J＝6.6Hz,2H),1.74(h,J＝7.1Hz,2H),0.97(t,J＝7.4Hz,3H).LC-MS(ESI)calcd for C11H12O3[M+H]+193.08,found 193.10.
(E)-4-(2-氯苯基)-2-(2-丙氧基甲酰基苄叉肼基)噻唑A2
Figure PCTCN2017084749-appb-000050
合成方法同A1，得到淡黄色固体粉末180mg，产率58％。1H NMR(400MHz, DMSO-d6,ppm):δ12.40(s,1H),8.71(s,1H),8.00(d,J＝7.9Hz,1H),7.88(d,J＝7.6,2.4Hz,2H),7.67(t,J＝7.6Hz,1H),7.52(q,J＝7.9Hz,2H),7.46-7.32(m,3H),4.26(t,J＝6.5Hz,2H),1.76(h,J＝7.1Hz,2H),0.98(t,J＝7.3Hz,3H).13C NMR(100MHz,DMSO-d6,ppm):δ167.68,167.03,147.63,140.16,134.95,133.6,132.71,131.53,131.22,130.85,130.59,129.52,129.4,129.38,127.73,126.74,109.35,67.06,21.96,10.87.HRMS(ESI)calcd for C20H18N3O2SCl[M+H]+400.0808,found400.0888.HPLCpurity:96.95％,Retention time＝12.44min.
2-甲酰基苯甲酸异丙酯3a
Figure PCTCN2017084749-appb-000051
合成方法同1a，得到黄色透明液体1.7g，产率88％。1H NMR(400MHz,DMSO-d6,ppm):δ10.40(s,1H),7.91-7.83(m,2H),7.80-7.75(m,2H),5.20(hept,J＝6.2Hz,1H),1.35(d,J＝6.2Hz,6H).LC-MS(ESI)calcd for C11H12O3[M+H]+193.08,found 193.10.
(E)-4-(2-氯苯基)-2-(2-异丙氧基甲酰基苄叉肼基)噻唑A3
Figure PCTCN2017084749-appb-000052
合成方法同A1，得到棕黄色固体粉末200mg，产率60％。1H NMR(400MHz,DMSO-d6,ppm):δ12.40(s,1H),8.69(s,1H),7.99(d,J＝7.8Hz,1H),7.90-7.81(m,2H),7.65(t,J＝7.6Hz,1H),7.55-7.46(m,2H),7.44-7.32(m,3H),5.16(hept,J＝6.3Hz,1H),1.35(d,J＝6.1Hz,6H).13C NMR(100MHz,DMSO-d6,ppm):δ205.55,167.21,166.04,147.14,139.59,134.28,133.20,132.07,131.02,130.72,130.35,130.07,129.35,129.00,128.84,127.21,126.11,108.82,68.85,21.53.HRMS(ESI)calcd for C20H18ClN3O2S[M+H]+400.0808,found 400.0868.HPLC purity:97.35％,Retention time＝10.44min.
2-甲酰基苯甲酸丁酯4a
Figure PCTCN2017084749-appb-000053
合成方法同1a，得到黄色澄清透明液体1.7g，产率86％。1H NMR(400MHz,DMSO-d6,ppm):δ10.41(s,1H),7.93-7.90(m,1H),7.88-7.85(m,1H),7.81-7.77 (m,2H),4.33(t,J＝6.5Hz,2H),1.71(p,J＝8.5,6.6Hz,2H),1.42(h,2H),0.94(t,J＝7.4Hz,3H).LC-MS(ESI)calcd for C12H14O3[M+H]+207.09,found 207.10.
(E)-4-(2-氯苯基)-2-(2-丁氧基甲酰基苄叉肼基)噻唑A4
Figure PCTCN2017084749-appb-000054
合成方法同A1，得到黄色粉末210mg，产率55％。1H NMR(400MHz,DMSO-d6,ppm):δ12.40(s,1H),8.71(s,1H),8.00(dd,J＝8.0,1.2Hz,1H),7.89-7.84(m,2H),7.67(td,J＝7.6,1.3Hz,1H),7.56-7.48(m,2H),7.42(td,J＝7.5,1.5Hz,1H),7.39-7.33(m,2H),4.30(t,J＝6.6Hz,2H),1.72(p,2H),1.42(h,2H),0.94(t,J＝7.4Hz,3H).13C NMR(100MHz,DMSO-d6,ppm):δ167.68,167.02,147.63,140.16,134.95,133.70,132.70,131.53,131.22,130.85,130.58,129.52,129.40,129.37,127.73,126.77,109.35,65.32,30.57,19.23,14.09.HRMS(ESI)calcd forC21H20N3O2SCl[M+H]+414.0965,found 414.1044.HPLC purity:97.15％,Retention time＝11.23min.
2-甲酰基苯甲酸-2-丁酯5a
Figure PCTCN2017084749-appb-000055
合成方法同1a，得到无色透明液体1.8g，产率91％。1H NMR(400MHz,DMSO-d6,ppm):δ10.39(s,1H),7.91-7.88(m,1H),7.87-7.84(m,1H),7.81-7.76(m,2H),5.06(q,J＝6.2Hz,1H),1.75-1.64(m,2H),1.32(d,J＝6.3Hz,3H),0.93(t,J＝7.4Hz,3H).LC-MS(ESI)calcd for C12H14O3[M+H]+207.09,found 207.10.
(E)-4-(2-氯苯基)-2-(2-(2-丁氧基)甲酰基苄叉肼基)噻唑A5
Figure PCTCN2017084749-appb-000056
合成方法同A1，得到浅黄色固体粉末150mg，产率52％。1H NMR(400MHz,DMSO-d6,ppm):δ12.41(s,1H),8.71(s,1H),8.00(d,J＝7.9,1.2Hz,1H),7.90-7.84(m,2H),7.69-7.64(m,1H),7.56-7.48(m,2H),7.42(td,J＝7.5,1.5Hz,1H), 7.39-7.33(m,2H),5.03(h,J＝6.3Hz,1H),1.77-1.63(m,2H),1.33(d,J＝6.3Hz,3H),0.93(t,J＝7.4Hz,3H).13C NMR(100MHz,DMSO-d6,ppm):δ167.71,166.60,147.64,140.09,134.89,133.70,132.64,131.53,131.22,130.86,130.52,129.74,129.52,129.39,127.73,126.67,109.35,73.65,28.66,19.65,10.07.HRMS(ESI)calcd for C21H20N3O2SCl[M+H]+414.0965,found 414.0988.HPLC purity:98.05％,Retention time＝11.14min.
2-甲酰基苯甲酸戊酯6a
Figure PCTCN2017084749-appb-000057
合成方法同1a，得到澄清液体1.6g，产率77％。1H NMR(400MHz,DMSO-d6,ppm):δ10.40(s,1H),7.93-7.89(m,1H),7.88-7.84(m,1H),7.81-7.77(m,2H),4.32(t,J＝6.6Hz,2H),1.72(p,J＝6.8Hz,2H),1.42-1.30(m,4H),0.92-0.87(m,3H).LC-MS(ESI)calcd for C13H16O3[M+H]+221.11,found 221.10.
(E)-4-(2-氯苯基)-2-(2-戊氧基甲酰基苄叉肼基)噻唑A6
Figure PCTCN2017084749-appb-000058
合成方法同A1，得到黄色粉末120mg，产率52％。1H NMR(400MHz,DMSO-d6,ppm):δ12.39(s,1H),8.71(s,1H),7.99(dd,J＝8.0,1.2Hz,1H),7.86(ddd,J＝7.8,6.1,1.6Hz,2H),7.66(td,J＝7.7,1.4Hz,1H),7.55-7.48(m,2H),7.42(td,J＝7.5,1.5Hz,1H),7.38-7.33(m,2H),4.29(t,J＝6.7Hz,2H),1.74(p,J＝6.8Hz,2H),1.42-1.30(m,4H),0.89(t,J＝7.0Hz,3H).13C NMR(100MHz,DMSO-d6,ppm):δ167.68,167.02,147.63,140.19,134.95,133.70,132.69,131.52,131.22,130.85,130.57,129.52,129.40,129.38,127.73,126.80,109.33,65.60,28.21,28.14,22.29,14.34.HRMS(ESI)calcd for C22H22N3O2SCl[M+H]+428.1121,found428.1201.HPLC purity:97.55％,Retention time＝9.44min.
2-甲酰基苯甲酸己酯7a
Figure PCTCN2017084749-appb-000059
合成方法同1a，得到无色透明液体1.9g，产率81％。1H NMR(400MHz, DMSO-d6,ppm):δ10.40(s,1H),7.93-7.88(m,1H),7.88-7.83(m,1H),7.82-7.76(m,2H),4.32(t,J＝6.6Hz,2H),1.71(p,2H),1.43-1.25(m,6H),0.90-0.83(m,3H).LC-MS(ESI)calcd for C14H18O3[M+H]+235.13,found 235.10.
(E)-4-(2-氯苯基)-2-(2-己氧基甲酰基苄叉肼基)噻唑A7
Figure PCTCN2017084749-appb-000060
合成方法同A1，得到淡黄色粉末110mg，产率59％。1H NMR(400MHz,DMSO-d6,ppm):δ12.45(s,1H),8.76(s,1H),8.05(d,J＝8.0Hz,1H),7.96-7.87(m,2H),7.72(t,J＝7.7Hz,1H),7.62-7.53(m,2H),7.51-7.37(m,3H),4.34(t,J＝6.6Hz,2H),1.79(t,J＝7.4Hz,2H),1.49-1.28(m,6H),0.92(t,J＝6.7Hz,3H).13C NMR(100MHz,DMSO-d6,ppm):167.56,166.92,147.53,140.10,134.85,133.60,132.56,131.41,131.12,130.74,130.44,129.39,129.30,129.27,127.60,126.71,109.20,65.51,31.27,28.36,25.52,22.36,14.22.HRMS(ESI)calcd for C23H24N3O2SCl[M+H]+442.1278,found 442.1355.HPLC purity:98.60％,Retention time＝13.41min.
2-甲酰基苯甲酸环己酯8a
Figure PCTCN2017084749-appb-000061
合成方法同1a，得到淡黄色透明液体1.8g，产率77％。1H NMR(400MHz,DMSO-d6,ppm):δ10.41(s,1H),7.93-7.88(m,1H),7.87-7.82(m,1H),7.81-7.75(m,2H),5.05-4.95(m,1H),1.97–1.88(m,2H),1.76-1.66(m,2H),1.61-1.47(m,3H),1.47-1.36(m,2H),1.36-1.25(m,1H).LC-MS(ESI)calcd for C14H16O3[M+H]+233.11,found 233.10.
(E)-4-(2-氯苯基)-2-(2-环己氧基甲酰基苄叉肼基)噻唑A8
Figure PCTCN2017084749-appb-000062
合成方法同A1，得到黄色粉末130mg，产率50％。1H NMR(400MHz,DMSO-d6,ppm):δ12.42(s,1H),8.70(s,1H),8.00(d,J＝7.8Hz,1H),7.90-7.83(m, 2H),7.66(t,J＝7.4Hz,1H),7.56-7.47(m,2H),7.42(td,J＝7.5,1.2Hz,1H),7.39-7.33(m,2H),5.00-4.91(m,1H),1.98-1.89(m,2H),1.78-1.68(m,2H),1.64-1.49(m,3H),1.47-1.25(m,3H).13C NMR(100MHz,DMSO-d6,ppm):δ167.72,166.37,140.10,134.84,132.62,131.52,131.22,130.86,130.60,129.80,129.52,129.39,127.73,126.67,109.34,73.85,31.44,25.34,23.69.HRMS(ESI)calcd for C23H22N3O2SCl[M+H]+440.1121,found 440.1101.HPLC purity:98.03％,Retention time＝13.03min.
2-甲酰基苯甲酸特戊酸甲基酯9a
Figure PCTCN2017084749-appb-000063
称取邻羧基苯甲醛(1g,6.7mmol)于100mL三口瓶中，加入碳酸铯(4.3g,13.3mmol)，碘化钾(1.1g,6.7mmol)，N,N-二甲基甲酰胺(DMF)15mL，电磁搅拌下加入特戊酸氯甲酯(1.5g,10mmol)，加热升温至70℃，反应20h，TLC检测至反应结束，停止加热，向反应瓶中加入20mL水，将反应液用二氯甲烷和水萃取三次，合并有机相，加无水硫酸钠干燥，抽滤，滤液加入等质量硅胶，过柱纯化，得到澄清透明液体0.9g，产率52％。LC-MS(ESI)calcd for C14H16O5[M+H]+265.08,found 265.10.
(E)-4-(2-氯苯基)-2-(2-特戊酸甲氧基甲酰基苄叉肼基)噻唑A9
Figure PCTCN2017084749-appb-000064
合成方法同A1，后处理得到粗品后，用反析法(粗品用适量乙酸乙酯溶解完全，电磁搅拌下加入石油醚，快速搅拌，析出固体，抽滤即可)得到淡黄色粉末180mg，产率49％。1H NMR(400MHz,DMSO-d6,ppm):δ12.43(s,1H),8.73(s,1H),8.03(d,J＝8.0,1.2Hz,1H),7.89-7.83(m,2H),7.74-7.68(m,1H),7.56-7.51(m,2H),7.42(td,J＝7.5,1.4Hz,1H),7.39-7.33(m,2H),5.98(s,2H),1.18(s,9H).13C NMR(100MHz,DMSO-d6,ppm):δ167.13,164.91,139.23,135.19,133.04,131.04,130.73,130.35,130.31,129.04,129.02,127.24,127.06,126.51,108.96,80.13,38.26,26.47.HRMS(ESI)calcd for C23H22N3O4SCl[M+H]+472.1020,found  472.1096.HPLC purity:97.79％,Retention time＝9.68min.
2-甲酰基苯甲酸异丙基碳酸甲基酯10a
Figure PCTCN2017084749-appb-000065
合成方法同9a，得到无色透明液体0.8g，产率45％。LC-MS(ESI)calcd for C13H14O6[M+NH4]+284.08,found 284.10.
(E)-4-(2-氯苯基)-2-(2-异丙基碳酸甲氧基甲酰基苄叉肼基)噻唑A10
Figure PCTCN2017084749-appb-000066
合成方法同A9，得到黄色粉末160mg，产率53％。1H NMR(400MHz,DMSO-d6,ppm):δ12.44(s,1H),8.76(s,1H),8.04(d,J＝8.0,1.2Hz,1H),7.93-7.85(m,2H),7.73(t,J＝7.6,1.3Hz,1H),7.57-7.51(m,2H),7.43(td,J＝7.5,1.5Hz,1H),7.40-7.33(m,2H),5.96(s,2H),4.85(hept,J＝6.2Hz,1H),1.26(d,J＝6.2Hz,6H).13C NMR(100MHz,DMSO-d6,ppm):δ167.62,165.32,153.25,147.67,139.76,135.83,133.66,131.54,131.23,131.01,130.85,129.54,129.49,127.74,127.30,126.99,109.47,83.02,73.24,21.80.HRMS(ESI)calcd for C22H20N3O5SCl[M+H]+474.0812,found 474.0889.HPLC purity:99.25％,Retention time＝12.74min.
2-甲酰基苯甲酸苯甲酯11a
Figure PCTCN2017084749-appb-000067
称取2-羧基苯甲醛(1.5g,10mmol)于100mL三口瓶B中，加入无水甲苯20mL，外部冰浴，电磁搅拌下加入氯化亚砜(1.79g,15mmol)，搅拌十五分钟后，撤去冰浴，将反应瓶B转移到油浴锅中，加热升温至110℃，回流反应4h，停止加热，旋干溶剂得到黄色粘稠液体，直接用10mL无水二氯甲烷稀释备用。另取100mL单口反应瓶A，向瓶中加入苯甲醇(1.08g,10mmol)，三乙胺(1.52g,15mmol)，无水二氯甲烷15mL，冰盐浴下电磁搅拌十五分钟，将酰氯的二氯甲 烷溶液逐滴加到反应瓶A中，冰浴反应三十分钟后，转室温反应，TLC检测反应，反应结束后，向反应液中加水搅拌十分钟，静置分层，取有机相，水相用二氯甲烷萃取三次，合并有机相，干燥，旋干溶剂，过柱纯化，得到无色透明液体1.1g，产率46％。1H NMR(400MHz,DMSO-d6,ppm):δ10.42(s,1H),7.95-7.91(m,2H),7.78-7.75(m,2H),7.50-7.48(m,2H),7.42-7.36(m,3H),5.39(s,2H).LC-MS(ESI)calcd for C15H12O3[M+H]+241.08,found 241.10.
(E)-4-(2-氯苯基)-2-(2-苯甲氧基甲酰基苄叉肼基)噻唑A11
Figure PCTCN2017084749-appb-000068
合成方法同A1，得到棕黄色粉末80mg，产率46％。1H NMR(400MHz,DMSO-d6,ppm):δ12.41(s,1H),8.78(s,1H),8.02(d,J＝7.9Hz,1H),7.92-7.86(m,2H),7.70-7.65(m,1H),7.56-7.49(m,4H),7.45-7.40(m,3H),7.39-7.34(m,3H),5.38(s,2H).13C NMR(100MHz,DMSO-d6,ppm):δ166.20,147.14,139.61,135.86,134.72,133.20,132.41,131.04,130.73,130.35,130.21,129.01,128.92,128.51,128.37,128.16,128.08,127.22,126.35,108.87,66.59.HRMS(ESI)calcd for C24H18N3O2SCl[M+H]+448.0808,found 448.0888.HPLC purity:98.45％,Retention time＝12.14min.
2-甲酰基苯甲酸苯乙酯12a
Figure PCTCN2017084749-appb-000069
合成方法同9a，得到透明液体2.5g，产率70％。1H NMR(400MHz,DMSO-d6,ppm):δ10.31(s,1H),7.86-7.81(m,2H),7.79-7.74(m,2H),7.32(d,J＝4.4Hz,4H),7.27-7.20(m,1H),4.56(t,J＝6.7Hz,2H),3.06(t,J＝6.7Hz,2H).LC-MS(ESI)calcd for C16H14O3[M+H]+255.09,found 255.10.
(E)-4-(2-氯苯基)-2-(2-苯乙氧基甲酰基苄叉肼基)噻唑A12
Figure PCTCN2017084749-appb-000070
合成方法同A1，得到黄色粉末100mg，产率51％。1H NMR(400MHz,DMSO-d6,ppm):δ12.46(s,1H),8.79(s,1H),8.05(d,J＝7.9Hz,1H),7.94(d,J＝7.7Hz,1H),7.84(d,J＝7.8Hz,1H),7.71(t,J＝7.7Hz,1H),7.60(d,J＝7.9Hz,1H),7.54(t,J＝7.6Hz,1H),7.48(t,J＝7.6Hz,1H),7.46-7.34(m,6H),7.32-7.25(m,1H),4.57(t,J＝6.8Hz,2H),3.14(t,J＝6.9Hz,2H).13C NMR(100MHz,DMSO-d6,ppm):δ167.58,166.74,147.55,140.03,138.30,134.93,133.61,132.66,131.43,131.13,130.75,130.45,129.41,129.24,129.03,128.78,127.62,126.80,126.67,109.25,65.98,34.61.HRMS(ESI)calcd for C25H20N3O2SCl[M+H]+462.0965,found 462.1041.HPLC purity:98.60％,Retention time＝12.24min.
2-甲酰基苯甲酰(N,N-二甲基)胺13a
Figure PCTCN2017084749-appb-000071
称取邻羧基苯甲醛(1.5g,10mmol)于100mL三口瓶中，加入无水二氯甲烷30mL，加入无水DMF四滴，外部冰盐浴，电磁搅拌下加入草酰氯(1.9g,15mmol)，搅拌三十分钟后，撤冰浴，转室温反应2h，检测反应完全，旋干溶剂得到黄色粘稠液体，直接用10mL无水二氯甲烷稀释备用。另取100mL单口反应瓶，向瓶中加入二甲胺(0.45g,10mmol)，三乙胺(1.52g,15mmol)，无水二氯甲烷10mL，冰浴下电磁搅拌十五分钟，将新制酰氯的二氯甲烷溶液经恒压滴液漏斗逐滴加到反应瓶中，冰浴反应三十分钟后，转室温反应，点板检测反应，反应结束后，向反应液中加水搅拌十分钟，静置分层，取有机相，水相用二氯甲烷萃取三次，合并有机相，干燥，旋干溶剂，过柱纯化，得到无色透明液体0.5g，产率26％。1H NMR(400MHz,DMSO-d6,ppm):δ9.97(s,1H),7.98-7.94(m,1H),7.75(td,J＝7.5,1.4Hz,1H),7.67-7.61(m,1H),7.44-7.39(m,1H),3.03(s,3H),2.72(s,3H).LC-MS(ESI)calcd for C10H11NO2[M+H]+178.08,found 178.10.
(E)-4-(2-氯苯基)-2-((N,N-二甲基)甲酰基苄叉肼基)噻唑A13
Figure PCTCN2017084749-appb-000072
合成方法同A1，得到黄色粉末80mg，产率31％。1H NMR(400MHz,DMSO-d6,ppm):δ12.25(s,1H),8.07(s,1H),7.94-7.89(m,2H),7.61-7.58(m,1H),7.57-7.53(m,1H),7.52-7.46(m,2H),7.44-7.40(m,2H),7.37-7.32(m,1H),3.13(s,3H),2.85(s,3H).13C NMR(100MHz,DMSO-d6,ppm):δ169.33,167.36,147.51,139.24,136.34,133.59,131.42,131.12,130.92,130.74,129.47,129.41,129.32,127.61,127.19,126.02,109.19,38.50,34.64.HRMS(ESI)calcd for C19H17N4OSCl[M+H]+385.0812,found 385.0802.HPLC purity:98.25％,Retention time＝11.45min.
2-甲基氨基硫脲4
Figure PCTCN2017084749-appb-000073
称取甲基肼硫酸盐(12.5g,86.5mmol)于500mL三口瓶中，加入350mL无水乙醇，电磁搅拌下加入硫氰酸铵(8g,104.0mmol)，升温至回流，反应过程中TLC检测，反应48h后，将反应停止，冷却至室温，抽滤，滤液旋干，加硅胶过柱层析(洗脱剂及比例：DCM:MeOH＝100:1v/v)，得白色粉末状固体2.2g，产率24％。1H NMR(400MHz,DMSO-d6,ppm):δ7.38(s,2H),4.88(s,2H),3.40(s,3H).LC-MS(ESI)calcd for C2H7N3S[M+H]+106.04,found 106.10.
2-甲基-1-(2-羧基苄叉基)氨基硫脲5
Figure PCTCN2017084749-appb-000074
称取化合物4(600mg,5.7mmol)于250mL三口瓶中，加入150mL无水乙醇，电磁搅拌下加入邻甲酰基苯甲酸(855mg,5.7mmol)，升温至回流，反应中用TLC监测反应，反应4h，反应结束，停止加热，将反应液冷却至室温，加硅胶旋干溶剂，过硅胶柱层析(洗脱剂及比例：DCM:MeOH＝100:1,v/v)，得到白色粉末状固体700mg，产率51.9％。1H NMR(400MHz,DMSO-d6,ppm):δ13.32(br,1H),8.54(s,1H),8.50(s,1H),8.33(d,J＝7.6Hz,1H),8.24(s,1H),7.90(d,J＝7.8Hz,1H),7.60(t,J＝7.2Hz,1H),7.52(t,J＝7.2Hz,1H),3.75(s,3H).LC-MS (ESI)calcd for C2H7N3S[M+H]+238.06,found 238.10.
(E)-4-(2-氯苯基)-2-[1-甲基-2-(2-羧基苄叉基)肼基]噻唑B
Figure PCTCN2017084749-appb-000075
称取化合物5(700mg,2.94mmol)于250mL三口瓶中，加入80mL无水乙醇，电磁搅拌下加入2-溴-2’-氯苯乙酮(0.46mL,2.94mmol)，加热至回流，反应用TLC检测，4h后，反应结束，停止加热将反应液冷却至室温，加硅胶后旋干溶剂，过硅胶柱层析(洗脱剂及比例：DCM:MeOH＝100:1,v/v)分离，得到黄色粉末状固体540mg，产率49％。Mp 210.2-210.8℃，1H NMR(400MHz,DMSO-d6,ppm):δ13.26(s,1H),8.60(s,1H),8.04(d,J＝7.6Hz,1H),7.97-7.89(m,2H),7.64(t,J＝7.8Hz,1H),7.55-7.48(m,2H),7.46(s,1H),7.40(td,J1＝7.6Hz,J2＝1.2Hz,1H),7.32(td,J1＝7.6Hz,J2＝1.6Hz,1H),3.65(s,3H).13C NMR(100MHz,DMSO-d6,ppm):δ168.96,168.50,147.22,136.91,135.21,133.40,132.46,131.50,131.02,130.88,130.55,130.14,129.46,129.18,127.62,126.50,111.43,32.90.HRMS(ESI)calcd for C18H15ClN3O2S[M+H]+372.0574,found 372.0573.HPLC purity:98.15％,Retention time＝9.46min.
(E)-4-(2-氯苯基)-2-[1-甲基-2-(2-乙氧基甲酰基苄叉基)肼基]噻唑B1
Figure PCTCN2017084749-appb-000076
称取化合物B(0.2g,0.54mmol)于50mL两口瓶中，加入碳酸铯(0.35g,1.08mmol)，N,N-二甲基甲酰胺(DMF)10mL，电磁搅拌下加入碘乙烷(0.18g,1.08mmol)，加热升温至70℃，反应过程中用TLC检测，直至反应5h后原料反应完全，停止加热，向反应瓶中加入20mL水，用二氯甲烷(DCM)萃取反应液，静置分层取有机相，水相用二氯甲烷萃取三次，合并有机相，干燥，旋干溶剂，过柱纯化，得到黄色粉末固体120mg，产率58％。1H NMR(400MHz,DMSO-d6,ppm):δ8.49(s,1H),8.02(d,J＝8.1,1.2Hz,1H),7.96(dd,J＝7.7,1.8Hz,1H),7.92-7.88(m,1H),7.69(t,J＝7.6,1.4Hz,1H),7.56-7.47(m,3H),7.45-7.40(m,1H),7.37(td,J＝7.6,1.8Hz,1H),4.36(q,J＝7.1Hz,2H),3.69(s,3H),1.35(t,J＝7.1Hz,3H).13C NMR(100MHz,DMSO-d6,ppm):δ168.51,166.58,146.72,136.19,134.55,133.00,132.32,131.12,130.69,130.36,130.36,129.08,129.06,128.88,127.25,126.36,111.10,61.16,32.55,14.04.HRMS(ESI)calcd for C20H18N3O2SCl [M+H]+400.0808,found 400.0886.HPLC purity:98.65％,Retention time＝16.44min.
(E)-4-(2-氯苯基)-2-[1-甲基-2-(2-丙氧基甲酰基苄叉基)肼基]噻唑B2
Figure PCTCN2017084749-appb-000077
合成方法同B1，得到黄色粉末固体100mg，产率51％。1H NMR(400MHz,DMSO-d6,ppm):δ8.49(s,1H),8.02(d,J＝8.0,1.2Hz,1H),7.96(dd,J＝7.7,1.9Hz,1H),7.91(dd,J＝7.9,1.4Hz,1H),7.72-7.67(m,1H),7.57-7.46(m,3H),7.43(td,J＝7.5,1.4Hz,1H),7.37(td,J＝7.6,1.8Hz,1H),4.27(t,J＝6.6Hz,2H),3.68(s,3H),1.75(h,J＝7.1Hz,2H),0.98(t,J＝7.4Hz,3H).13C NMR(100MHz,DMSO-d6,ppm):δ169.00,167.14,147.22,136.65,135.11,133.49,132.84,131.62,131.19,130.87,130.81,129.59,129.53,129.42,127.76,126.94,111.61,67.09,33.05,21.99,10.89.HRMS(ESI)calcd for C21H20N3O2SCl[M+H]+414.0965,found 414.1042.HPLC purity:99.75％,Retention time＝15.44min.
(E)-4-(2-氯苯基)-2-[1-甲基-2-(2-异丙氧基甲酰基苄叉基)肼基]噻唑B3
Figure PCTCN2017084749-appb-000078
合成方法同B1，得到淡黄色粉末140mg，产率57％。1H NMR(400MHz,DMSO-d6,ppm):δ8.48(s,1H),8.02(d,J＝8.0,1.2Hz,1H),7.96(dd,J＝7.7,1.8Hz,1H),7.88(d,J＝7.8,1.4Hz,1H),7.71-7.65(m,1H),7.56-7.47(m,3H),7.46-7.39(m,1H),7.36(td,J＝7.6,1.8Hz,1H),5.21(hept,J＝6.3Hz,1H),3.69(s,3H),1.36(d,J＝6.2Hz,6H).13C NMR(100MHz,DMSO-d6,ppm):δ168.52,166.09,146.73,136.11,134.45,133.01,132.24,131.12,130.70,130.36,129.43,129.08,128.87,127.25,126.27,111.10,68.75,32.54,21.59.HRMS(ESI)calcd for C21H20N3O2SCl[M+H]+414.0965,found 414.0955.HPLC purity:99.65％,Retention time＝12.44min.
(E)-4-(2-氯苯基)-2-[1-甲基-2-(2-丁氧基甲酰基苄叉基)肼基]噻唑B4
Figure PCTCN2017084749-appb-000079
合成方法同B1，得到黄色粉末固体190mg，产率61％。1H NMR(400MHz,DMSO-d6,ppm):δ8.49(s,1H),8.01(d,J＝7.9,1.2Hz,1H),7.96(dd,J＝7.7,1.8Hz,1H),7.90(dd,J＝7.8,1.3Hz,1H),7.72-7.66(m,1H),7.56-7.46(m,3H),7.43(td,J＝7.5,1.4Hz,1H),7.36(td,J＝7.6,1.9Hz,1H),4.31(t,J＝6.6Hz,2H),3.68(s,3H),1.71(p,2H),1.42(h,2H),0.93(t,J＝7.4Hz,3H).13C NMR(100MHz,DMSO-d6,ppm):δ168.98,167.14,147.21,136.62,135.11,133.49,132.81,131.62,131.18,130.86,130.80,129.58,129.50,129.40,127.75,126.95,111.60,65.33,33.03,30.60,19.26,14.06.HRMS(ESI)calcd for C22H22N3O2SCl[M+H]+428.1121,found 428.1131.HPLC purity:98.65％,Retention time＝20.44min.
(E)-4-(2-氯苯基)-2-[1-甲基-2-(2-(2-丁氧基)甲酰基苄叉基)肼基]噻唑B5
Figure PCTCN2017084749-appb-000080
合成方法同B1，得到淡黄色粉末160mg，产率56％。1H NMR(400MHz,DMSO-d6,ppm):δ8.49(s,1H),8.02(dd,J＝8.0,1.2Hz,1H),7.96(dd,J＝7.7,1.8Hz,1H),7.90(dd,J＝7.8,1.4Hz,1H),7.72-7.66(m,1H),7.56-7.46(m,3H),7.43(td,J＝7.5,1.4Hz,1H),7.37(td,J＝7.6,1.9Hz,1H),5.06(h,J＝6.3Hz,1H),3.69(s,3H),1.76-1.62(m,2H),1.33(d,J＝6.3Hz,3H),0.93(t,J＝7.5Hz,3H).13C NMR(100MHz,DMSO-d6,ppm):δ169.02,166.68,147.22,136.57,135.04,133.49,132.80,131.62,131.19,130.87,130.79,129.81,129.60,129.43,127.76,126.84,111.62,73.57,33.04,28.71,19.66,10.04.HRMS(ESI)calcd for C22H22N3O2SCl[M+H]+428.1121,found 428.1131.HPLC purity:98.95％,Retention time＝18.44min.
(E)-4-(2-氯苯基)-2-[1-甲基-2-(2-戊氧基甲酰基苄叉基)肼基]噻唑B6
Figure PCTCN2017084749-appb-000081
合成方法同B1，得到淡黄色粉末130mg，产率55％。1H NMR(400MHz,DMSO-d6,ppm):δ8.48(s,1H),8.04-7.85(m,3H),7.69(t,J＝7.6Hz,1H),7.59- 7.31(m,5H),4.30(t,J＝6.5Hz,2H),3.68(s,3H),1.78-1.64(m,2H),1.35(s,4H),0.88(t,J＝6.7Hz,3H).13C NMR(100MHz,DMSO-d6,ppm):δ168.99,167.18,147.22,136.69,135.08,133.50,132.80,131.62,131.19,130.87,130.77,129.60,129.57,129.42,127.76,127.01,111.60,65.64,33.05,28.26,28.19,22.28,14.33.HRMS(ESI)calcd for C23H24N3O2SCl[M+H]+442.1278,found 442.1265.HPLC purity:98.21％,Retention time＝22.10min.
(E)-4-(2-氯苯基)-2-[1-甲基-2-(2-己氧基甲酰基苄叉基)肼基]噻唑B7
Figure PCTCN2017084749-appb-000082
合成方法同B1，得到浅黄色粉末170mg，产率49％。1H NMR(400MHz,DMSO-d6,ppm):δ8.48(s,1H),8.02-7.94(m,2H),7.88(dd,J＝7.8,1.3Hz,1H),7.72-7.66(m,1H),7.58-7.46(m,3H),7.43(td,J＝7.6,1.4Hz,1H),7.36(td,J＝7.6,1.9Hz,1H),4.30(t,J＝6.6Hz,2H),3.68(s,3H),1.75-1.66(m,2H),1.41-1.25(m,6H),0.88-0.82(m,3H).13C NMR(100MHz,DMSO-d6,ppm):δ237.52,235.49,158.25,145.81,132.69,131.51,131.35,131.07,130.76,129.48,129.31,127.65,126.94,125.32,124.37,91.91,65.55,32.94,31.27,28.41,25.58,22.38,14.24.HRMS(ESI)calcd for C24H26N3O2SCl[M+H]+456.1434,found 456.1410.HPLC purity:97.55％,Retention time＝17.44min.
(E)-4-(2-氯苯基)-2-[1-甲基-2-(2-环己氧基甲酰基苄叉基)肼基]噻唑B8
Figure PCTCN2017084749-appb-000083
合成方法同B1，得到淡黄色粉末130mg，产率53％。1H NMR(100MHz,DMSO-d6,ppm):δ8.50(s,1H),8.02(d,J＝8.0,1.3Hz,1H),7.96(dd,J＝7.7,1.8Hz,1H),7.90(dd,J＝7.9,1.4Hz,1H),7.72-7.66(m,1H),7.56-7.47(m,3H),7.43(td,J＝7.5,1.5Hz,1H),7.37(td,J＝7.6,1.9Hz,1H),5.03-4.95(m,1H),3.69(s,3H),1.98-1.90(m,2H),1.79–1.65(m,2H),1.61-1.49(m,3H),1.47-1.29(m,3H).13C NMR(100MHz,DMSO-d6,ppm):δ169.02,166.44,147.22,136.65,135.03,133.50,132.81,131.62,131.20,130.87,129.89,129.61,129.43,127.77,126.82,111.62,73.78,33.06,31.51,25.32,23.69.HRMS(ESI)calcd for C24H24N3O2SCl [M+H]+454.1278,found 454.1294.HPLC purity:97.32％,Retention time＝11.53min.
(E)-4-(2-氯苯基)-2-[1-甲基-2-(2-苯乙氧基甲酰基苄叉基)肼基]噻唑B9
Figure PCTCN2017084749-appb-000084
合成方法同B1，不同之处为加入等摩尔量的碘化钾，得到黄色粉末100mg，产率46％。1H NMR(400MHz,DMSO-d6,ppm):δ8.40(s,1H),8.02-7.93(m,2H),7.84-7.80(m,1H),7.71-7.66(m,1H),7.56-7.47(m,3H),7.43(td,J＝7.5,1.5Hz,1H),7.37(td,J＝7.6,1.9Hz,1H),7.31(d,J＝4.3Hz,4H),7.26-7.19(m,1H),4.54(t,J＝6.7Hz,2H),3.61(s,3H),3.06(t,J＝6.7Hz,2H).13C NMR(100MHz,DMSO-d6,ppm):δ168.88,166.89,147.13,138.34,136.45,135.03,133.42,132.77,131.52,131.10,130.76,130.60,129.49,129.24,128.77,127.65,126.90,126.80,111.49,65.95,34.67,32.92.HRMS(ESI)calcd for C26H22N3O2SCl[M+H]+476.1121,found 476.1201.HPLC purity:98.35％,Retention time＝16.44min.
(E)-4-(2-氯苯基)-2-[1-甲基-2-(2-特戊酸甲氧基甲酰基苄叉基)肼基]噻唑B10
Figure PCTCN2017084749-appb-000085
合成方法同B9，得到亮黄色粉末120mg，产率57％。1H NMR(400MHz,DMSO-d6,ppm):δ8.44(s,1H),8.03(d,J＝8.0,1.2Hz,1H),7.96(dd,J＝7.7,1.9Hz,1H),7.89-7.85(m,1H),7.77-7.71(m,1H),7.57-7.53(m,2H),7.49(s,1H),7.43(td,J＝7.5,1.4Hz,1H),7.37(td,J＝7.6,1.9Hz,1H),5.99(s,2H),3.69(s,3H),1.17(s,9H).13C NMR(100MHz,DMSO-d6,ppm):δ176.91,168.94,165.74,147.24,136.17,135.56,133.60,133.48,131.64,131.20,130.97,130.87,129.62,129.56,128.04,127.77,127.18,111.73,89.14,80.79,33.07,26.96.HRMS(ESI)calcd for C24H24N3O4SCl[M+H]+486.1176,found 486.1256.HPLC purity:97.64％,Retention time＝13.15min.
(E)-4-(2-氯苯基)-2-[1-甲基-2-(2-异丙基碳酸甲氧基甲酰基苄叉基)肼基]噻唑B11
Figure PCTCN2017084749-appb-000086
合成方法同B9，得到萤黄色粉末90mg，产率54％。1H NMR(400MHz,DMSO-d6,ppm):δ8.74(s,1H),8.30(d,J＝8.0,1.3Hz,1H),8.15(dd,J＝8.0,1.4Hz,1H),8.09(dd,J＝7.8,1.8Hz,1H),7.74-7.67(m,1H),7.54-7.47(m,2H),7.44-7.36(m,2H),7.36-7.28(m,2H),6.10(s,2H),5.07-5.00(m,1H),3.86(s,3H),1.42(d,J＝6.2Hz,6H).13C NMR(100MHz,DMSO-d6,ppm):δ169.03,165.22,153.36,147.30,136.67,134.75,133.43,133.13,131.72,131.21,131.12,130.39,128.36,128.31,126.86,126.71,126.38,110.68,82.11,73.18,32.67,21.57.HRMS(ESI)calcd for C23H22N3O5SCl[M+H]+488.0969,found 488.1046.HPLC purity:96.65％,Retention time＝17.44min.
(E)-4-(2-氯苯基)-2-[1-甲基-2-(2-(氯甲酰基苄叉基)肼基]噻唑12b
Figure PCTCN2017084749-appb-000087
称取化合物B(0.2g,0.54mmol)于50mL两口瓶中，加入二氯甲烷10mL，N,N-二甲基甲酰胺(DMF)2d，冰浴搅拌十分钟，取草酰氯(0.13g,1.08mmol)溶于5mL二氯甲烷中逐滴滴加入反应瓶中。继续冰浴，半小时，撤冰浴，转室温反应2h，检测反应：取反应液于小样品管内加甲醇震荡后加水和乙酸乙酯，静置分层，取上层有机相点板，发现原料点基本消失。说明酰氯制备成功。将反应液溶剂旋干，得到黄色液体。直接用于下一步反应。
(E)-4-(2-氯苯基)-2-[1-甲基-2-(2-((N，N-二甲基)甲酰基苄叉基)肼基]噻唑B12
Figure PCTCN2017084749-appb-000088
取50mL单口反应瓶，向瓶中加入二甲胺(24mg,0.54mmol)，三乙胺(55mg,0.54mmol)，无水二氯甲烷10mL，冰浴下电磁搅拌十五分钟，将新制的酰氯二氯甲烷溶液逐滴加到反应瓶中，冰浴反应三十分钟后，转室温反应10h，用TLC检测反应，反应结束后，向反应液中加水和二氯甲烷各5mL，搅拌十分钟，静置分层，取有机相，水相用二氯甲烷萃取三次，合并有机相，干燥，旋干溶剂， 过柱纯化，得到灰白的固体粉末50mg，产率25％。1H NMR(400MHz,DMSO-d6,ppm):δ7.96(dd,J＝8.0,2Hz,1H),7.87(d,J＝7.6Hz,1H),7.73(s,1H),7.55-7.29(m,7H),3.65(s,3H),3.07(s,3H),2.80(s,3H).13C NMR(100MHz,DMSO-d6,ppm):δ169.57,168.91,147.14,136.30,135.65,133.52,131.62,131.34,131.19,130.86,129.58,129.46,127.76,127.59,127.22,111.59,38.62,34.90,32.95.HRMS(ESI)calcd for C20H19N4OSCl[M+H]+399.0968,found 399.1048.HPLC purity:96.75％,Retention time＝11.14min.
实施例2(活性评价)
1.实验材料
1.1实验动物
SPF级BALB/c雄性小鼠，每只体重18～22g，购自上海斯莱克实验动物有限公司，适应性喂养7天后用于实验。
1.2试剂
2，4，6-三硝基苯磺酸(TNBS)购于美国Sigma公司，柳氮磺胺嘧啶片(SASP，0.25g/片)购于上海三维制药有限公司，无水乙醇购于上海泰坦科技股份有限公司，水合氯醛购于江苏强盛功能化学股份有限公司，氯化钠购于国药集团化学试剂有限公司，37％～40％甲醛溶液购于上海凌峰化学试剂有限公司，羧甲基纤维素钠购于上海晶纯生化科技股份有限公司，石蜡购于德国Leica公司，二甲苯购于上海凌峰化学试剂有限公司，苏木素伊红(H&E)染色试剂盒购于碧云天生物技术研究所，盐酸购于上海凌峰化学试剂有限公司，中性树胶购于国药集团化学试剂有限公司。
1.3主要实验仪器
电子天平购于上海精天电子仪器有限公司，精密电子天平购于梅特勒-托利多公司，游标卡尺购于哈尔滨量具刃具集团有限责任公司，显微镜购于Nikon公司，石蜡切片机购于德国Leica公司，电热恒温鼓风干燥箱购于上海一恒科学仪器有限公司。
2.实验方法
2.1实验动物饲养
BALB/c雄性小鼠饲养在温度20～25℃，相对湿度50～60％，白光、黑夜各12h的动物房中，可自由索食和饮水。
2.2实验动物分组
实验动物分组及给药剂量见表1。将小鼠随机分为正常对照组、三硝基苯磺酸(TNBS)模型组、药物治疗组及阳性药对照组(柳氮磺胺嘧啶500mg/kg)。
将小鼠随机分为四组，分组及每日给药情况如表1所示：
表1：小鼠分组及每日给药情况
Figure PCTCN2017084749-appb-000089
2.3建立TNBS诱导的小鼠溃疡性结肠炎模型与给药
所有模型及给药组小鼠腹部剃毛(面积2×2cm)，用1％TNBS(溶于50％乙醇)涂抹致敏。第7天小鼠禁食不禁水，24h后，腹腔注射0.1ml10％水合氯醛进行麻醉，2％TNBS灌肠，建立溃疡性结肠炎小鼠模型。将大鼠灌胃针插入小鼠结肠，针头尖端距肛门约3～3.5cm，注入0.1ml造模溶液(5％TNBS与去离子水按体积比4:1混合，所得溶液再与无水乙醇按体积比1:1混合所得)。仰卧放回鼠笼，自然苏醒。正常对照组以同样的方法注入0.12ml溶剂(50％乙醇溶液)。建模24h后随机处死2只造模小鼠，观察结肠的变化，以确认造模是否成功。
建模当天开始给药(用0.5％羧甲基纤维素钠，CMC-Na)，连续给药7天。每天称量体重，观察小鼠的精神状态、大便性状、皮毛状态、摄食量等。
2.4样品采集
最后一次给药24h后，颈部脱臼处死小鼠，剪开腹膜，取全部结肠，沿肠系膜缘纵向剖开肠管，用冰生理盐水冲洗干净，放在冰上，观察结肠的病理变化，然后除去脂肪和肠系膜，用滤纸吸干水分，称量结肠的重量并测量其长度。放入4％甲醛中固定24h以上，组织脱水，进行石蜡包埋、H&E染色，对病理切片观察评价。
2.5评价指标
2.5.1一般情况观察
每日观察并记录小鼠的精神状态、皮毛色泽以及活动情况等。每日称量小鼠体重，观察大便性状，根据疾病活动指数(DAI)体重评分标准如下：
体重：体重不变，0分；体重比正常下降1％～5％，1分；下降6％～10％，2分；11％～15％，3分；大于15％，4分。
2.5.2结肠重量/长度比值变化
取出小鼠结肠后，用精密电子天平测量结肠的重量，用游标卡尺测量其长度，计算结肠重量/长度的比值。
2.5.3结肠大体形态损伤情况观察
取结肠后，观察结肠与其他组织黏连、溃疡和炎症情况，并记录评分，评分标准为：无黏连、无溃疡、无炎症，0分；轻度黏连、局部充血、无溃疡，1分；重度黏 连、有1处溃疡(面积小于1cm)、无明显炎症，2分；1处以上溃疡(面积小于1cm)伴有炎症，3分；2处严重溃疡(面积大于1cm)并伴有炎症，4分；溃疡面积每增加1cm加1分。
2.5.4结肠组织病理学观察
光镜下观察结肠组织病理学改变情况，记录评分。评分标准如下：
1)上皮细胞：正常形态，0分；有杯状细胞丢失，1分；杯状细胞大面积丢失，2分；隐窝细胞丢失，3分；隐窝细胞大面积丢失，4分。
2)炎症细胞浸润：没有浸润，0分；浸润在隐窝基底层，1分；浸润到达黏膜肌层，2分；浸润深入到黏膜肌层，伴随黏膜增厚和明显水肿，3分；浸润到达黏膜下层，4分。
2.6统计学分析
数据以均值±标准差(mean±S.E.M.)表示，使用SPSS统计软件进行数据分析，组间比较应用one-way ANOVA和Tukey’s HSD post hoc检验，p<0.05认为有统计学意义。
3.实验结果和分析
3.1一般情况观察结果
跟正常对照组相比，TNBS模型组小鼠在造模第3天大便性状开始发生变化，出现腹泻症状，第4天后腹泻，血便，精神萎靡，活动减少，饮食减退，皮毛凌乱暗哑，体重明显下降；药物治疗组和阳性药对照组，在造模第3天出现腹泻症状，第4天后腹泻，血便，精神萎靡，活动减少，饮食减退，皮毛凌乱暗哑，体重下降，但跟TNBS模型组相比，症状较轻。体重下降情况如表2所示：
表2：小鼠体重下降情况
Figure PCTCN2017084749-appb-000090
Figure PCTCN2017084749-appb-000091
注：与正常对照组相比，**p<0.01：与模型组相比，#p<0.05,##p<0.01
3.2结肠重量/长度比值变化
结肠重量、长度以及二者比值如表3所示：
表3：小鼠重量/长度比值变化
Figure PCTCN2017084749-appb-000092
注：与正常对照组相比，*p<0.05；与模型组比，#p<0.05
TNBS模型组重量/长度比值明显比正常对照组大，表明模型组充血肿胀情况比较严重，而药物治疗组和阳性药对照组重量/长度比值明显低于模型组，表明本发明化合物和阳性药柳氮磺胺嘧啶都可以逆转TNBS引起的小鼠结肠充血水肿。
3.3结肠大体形态损伤情况与结肠组织病理学观察结果
观察结肠大体形态和组织病理学切片可以发现，跟正常对照组相比，模型组损伤比较明显，充血肿胀、溃疡严重，有炎症细胞的浸润现象。药物治疗组和阳性对照组，这些损伤情况明显得到改善(如图1所示)。
结肠大体形态损伤与组织病理学评分结果总结于表4。
表4：结肠大体形态损伤与组织病理学观察结果
Figure PCTCN2017084749-appb-000093
Figure PCTCN2017084749-appb-000094
注：与正常对照组相比，**p<0.01；与TNBS模型组相比，#p<0.05，##p<0.01
讨论：
从在动物水平进行的本发明化合物的溃疡性结肠炎治疗活性检测结果可知，本发明的化合物具备优异的溃疡性结肠炎治疗活性，其中一些化合物的溃疡性结肠炎治疗活性与阳性对照药物相当。从而为开发低毒、高效、安全的新型溃疡性结肠炎治疗药物奠定新的物质基础。
在本发明提及的所有文献都在本申请中引用作为参考，就如同每一篇文献被单独引用作为参考那样。此外应理解，在阅读了本发明的上述讲授内容之后，本领域技术人员可以对本发明作各种改动或修改，这些等价形式同样落于本申请所附权利要求书所限定的范围。

Claims (10)

  1. 式I所示化合物或其药学上可接受的盐或酯在制备治疗炎性肠病的药物中的用途：
    Figure PCTCN2017084749-appb-100001
    式中，
    R1选自：H、取代或未取代的C1-C6烷基、C3-C6环烷基；独立
    R2独立选自：H、卤素、取代或未取代的C1-C6烷基、取代或未取代的C1-C6烷氧基、CN、NO2、羟基、NRaRb；
    Ra、Rb可独立选自H或C1-C6烷基
    R3选自：H、取代或未取代的C1-C6烷基；
    R4选自：H、卤素；
    m为0～4的整数；
    n为0～5的整数。
  2. 如权利要求1所述的用途，其特征在于，所述炎性肠病是溃疡性结肠炎或克罗恩病；优选溃疡性结肠炎。
  3. 如权利要求1或2所述的用途，其特征在于，在式I中，
    R1选自：H、取代或未取代的C1-C6烷基；
    R2选自：H、卤素、取代或未取代的C1-C6烷基、取代或未取代的C1-C6烷氧基；
    R3选自：H、取代或未取代的C1-C6烷基；
    R4选自：H、卤素；
    m为0～2的整数；
    n为0～2的整数。
  4. 如权利要求3所述的用途，其特征在于，所述化合物如式II所示：
    Figure PCTCN2017084749-appb-100002
    式中，
    R1选自：H、取代或未取代的C1-C3烷基；
    R5和R6独立选自：H、卤素、取代或未取代的C1-C3烷基；
    R3选自：H、取代或未取代的C1-C3烷基；
    R7、R8和R9独立选自：H、卤素。
  5. 如权利要求1或2所述的用途，其特征在于，式I所示化合物或其药学上可接受的盐或酯是选自下组的化合物或其药学上可接受的盐或酯：
    Figure PCTCN2017084749-appb-100003
    Figure PCTCN2017084749-appb-100004
    Figure PCTCN2017084749-appb-100005
    Figure PCTCN2017084749-appb-100006
  6. 如权利要求5所述的用途，其特征在于，所述化合物是选自下组的化合物：
    Figure PCTCN2017084749-appb-100007
    Figure PCTCN2017084749-appb-100008
    Figure PCTCN2017084749-appb-100009
  7. 如权利要求6所述的用途，其特征在于，所述化合物是选自下组的化合物：
    Figure PCTCN2017084749-appb-100010
    Figure PCTCN2017084749-appb-100011
    Figure PCTCN2017084749-appb-100012
    Figure PCTCN2017084749-appb-100013
  8. 一种药物组合物在制备用于治疗炎性肠病的药物中的用途，其特征在于，所述药物组合物含有权利要求1-7中任一项所述的化合物或其药学上可接受的盐或酯，以及药学上可接受的载体或赋形剂。
  9. 一种药物组合物，其特征在于，所述药物组合物用于治疗炎性肠病并且含有权利要求1-7中任一项所述的化合物或其药学上可接受的盐或酯，以及药学上可接受的载体或赋形剂。
  10. 一种治疗炎性肠病的方法，所述方法包括将权利要求1-7中任一项所述的化合物或权利要求9所述的药物组合物给予需要治疗炎性肠病的对象。
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Citations (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US4319026A (en) * 1980-08-14 1982-03-09 Gulf Oil Corporation Heterocyclic-substituted hydrazides and hydrazones as plant growth regulators

Family Cites Families (7)

* Cited by examiner, † Cited by third party
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CN103127095A (zh) * 2011-11-23 2013-06-05 华东理工大学 二氢噻吩酮衍生物作为pfDHODH抑制剂的合成及应用
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Patent Citations (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US4319026A (en) * 1980-08-14 1982-03-09 Gulf Oil Corporation Heterocyclic-substituted hydrazides and hydrazones as plant growth regulators

Non-Patent Citations (1)

* Cited by examiner, † Cited by third party
Title
LI, SHILIANG ET AL.: "Rational Design of benzylidenehydrazinyl-substituted Thiazole Derivatives as Potent Inhibitors of Human Dihydroorotate Dehydrogenase with in Vivo Anti-arthritic Activity", SCIENTIFIC REPORTS, vol. 5, 7 October 2015 (2015-10-07), XP055439180 *

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