Pyrazole derivative and use thereof

Abstract

Provided are a compound of formula (I) or formula (II), or a pharmaceutically acceptable salt, cocrystal, stereoisomer, solvate, prodrug or metabolite thereof, and the use thereof in treating obesity or obesity complications.

Description

Pyrazole derivatives and uses thereof Technical Field

[0001] The present application relates to the field of medicine, and in particular to a pyrazole derivative and its use in treating or preventing obesity or obesity complications. Background Art

[0002] The endocannabinoid system (ECS) consists of cannabinoid receptors (CB1 and CB2), their ligands (anandamide and 2-AG), and enzymes that regulate their synthesis and degradation (monoacylglycerol lipase and fatty acid amide hydrolase). The ECS plays an important role in regulating a variety of physiological functions, including food intake, energy metabolism, emotional behavior, pain, cell division, and inflammation.

[0003] Both CB1 and CB2 belong to the G protein-coupled receptor superfamily. CB1 receptors are widely expressed in central tissues such as the cortex, hippocampus, amygdala, pituitary gland, and hypothalamus, and are also distributed in peripheral organs and tissues including the thyroid, adrenal glands, reproductive organs, bone, fat, liver, muscle, pancreas, kidneys, and gastrointestinal tract. CB1 receptors bind to cannabinoids and their derivatives, activating intracellular signaling and mediating a wide range of biological functions in the ECS, such as increasing appetite, promoting lipogenesis, inducing insulin resistance, and triggering dyslipidemia. Studies have shown that overactivation of CB1 receptors is closely associated with the development of visceral obesity, type 2 diabetes mellitus (T2DM), and its related complications. Therefore, inhibiting overactivation of CB1 is considered a potential treatment for obesity, T2DM, and non-alcoholic fatty liver disease (NAFLD). Rimonabant is the world's first CB1 inhibitor-based weight loss drug and was approved for marketing in the European Union in 2006. Later studies demonstrated an increased risk of severe psychiatric adverse reactions, including depression, anxiety, and suicide, leading to its global withdrawal from the market in October 2008. Rimonabant's central nervous system side effects are due to its ability to penetrate the blood-brain barrier and block CB1 receptors in the brain. Therefore, the development of highly peripherally selective CB1 receptor antagonists would reduce or eliminate these central nervous system side effects, providing a safer and more effective medication option for the prevention or treatment of conditions such as obesity, diabetes, dyslipidemia, and cardiovascular disease. Summary of the Invention

[0004] The object of the present invention is to provide a CB1 inhibitor, a pharmaceutical composition comprising the same, and use thereof in treating or preventing obesity or obesity complications.

[0005] In one aspect, the present invention relates to a compound of formula (I), or a pharmaceutically acceptable salt, cocrystal, stereoisomer, solvate, prodrug or metabolite thereof:

[0006] in,

[0007] Ring A represents a five-membered or six-membered monocyclic heteroaryl group containing 1 or 2 heteroatoms independently selected from N and S, preferably selected from thienyl, pyridinyl and thiazolyl, more preferably selected from thien-2-yl, pyridin-2-yl and thiazol-5-yl;

[0008] R1 is C 1-6 Halogenated alkyl, preferably C 1-3 Haloalkyl, more preferably C 1-3 Fluoroalkyl, most preferably trifluoromethyl;

[0009] m is 0, 1 or 2, R2 is independently selected from halogen, cyano, and C 1-6 Halogenated alkyl groups are preferably each independently selected from fluorine, chlorine, cyano, and C 1-3 Haloalkyl (especially C 1-3 Fluoroalkyl, more particularly trifluoromethyl), m is more preferably 1 or 2, particularly preferably 1;

[0010] R 3a 、R 3b 、R 4a and R 4b are each independently selected from hydrogen and C 1-6 Alkyl groups are preferably each independently selected from hydrogen and C 1-3 Alkyl, more preferably each independently selected from hydrogen and methyl, most preferably R 3a and R 3b At least one of them is hydrogen, and R 4a and R 4b Each is hydrogen.

[0011] Preferably, the compound of formula (I) is selected from:

[0012] In yet another aspect, the present invention relates to a compound of formula (II), or a pharmaceutically acceptable salt, cocrystal, stereoisomer, solvate, prodrug or metabolite thereof:

[0013] in,

[0014] Ring B represents a phenyl group or a five-membered or six-membered monocyclic heteroaryl group containing 1 or 2 heteroatoms independently selected from N and S, preferably selected from phenyl, thienyl, and pyridinyl, more preferably selected from phenyl, thien-2-yl, and pyridin-2-yl;

[0015] Ring C represents phenyl or thienyl, more preferably selected from phenyl and thien-2-yl;

[0016] R1 is C 1-6 Halogenated alkyl, preferably C 1-3Haloalkyl, more preferably C 1-3 Fluoroalkyl, most preferably trifluoromethyl;

[0017] m is 0, 1 or 2, R2 is independently selected from halogen, cyano, and C 1-6 Halogenated alkyl groups are preferably each independently selected from fluorine, chlorine, cyano, and C 1-3 Haloalkyl (especially C 1-3 Fluoroalkyl, more particularly trifluoromethyl), m is more preferably 1 or 2, particularly preferably 1;

[0018] R 3a 、R 3b 、R 4a and R 4b are each independently selected from hydrogen and C 1-6 Alkyl groups are preferably each independently selected from hydrogen and C 1-3 Alkyl, more preferably each independently selected from hydrogen and methyl, most preferably R 3a and R 3b At least one of them is hydrogen, and R 4a and R 4b Each is hydrogen.

[0019] Preferably, the compound of formula (II) is selected from:

[0020] In yet another aspect, the present invention relates to a compound, or a pharmaceutically acceptable salt, cocrystal, stereoisomer, solvate, prodrug or metabolite thereof, wherein the compound is selected from:

[0021] On the other hand, the present invention also relates to a pharmaceutical composition comprising:

[0022] (1) The compound as described above, or a pharmaceutically acceptable salt, cocrystal, stereoisomer, solvate, prodrug or metabolite thereof;

[0023] (2) optionally one or more other active ingredients; and

[0024] (3) Pharmaceutically acceptable carriers and / or excipients.

[0025] In another aspect, the present invention also relates to the use of the compound as described above, or a pharmaceutically acceptable salt, cocrystal, stereoisomer, solvate, prodrug or metabolite thereof, in the preparation of a medicament for treating or preventing obesity or obesity complications.

[0026] In a preferred embodiment, the complications of obesity are selected from one or more of diabetes, dyslipidemia, metabolic syndrome, dementia, inflammatory disease, cardiovascular disease, liver disease, or cancer.

[0027] Further preferably, diabetes is selected from one or more of type 1 diabetes, type 2 diabetes, impaired glucose tolerance or insulin resistance; dyslipidemia is selected from one or more of poor blood lipid levels, low levels of high-density lipoprotein, high levels of low-density lipoprotein or high levels of triglycerides; inflammatory diseases are selected from one or more of osteoarthritis, rheumatoid arthritis, inflammatory bowel disease or obesity-related inflammation; cardiovascular diseases are selected from one or more of atherosclerosis, hypertension, stroke or heart attack; liver disease is selected from one or more of liver inflammation, liver fibrosis, non-alcoholic steatohepatitis, fatty liver, liver enlargement, alcoholic liver disease, jaundice, cirrhosis or hepatitis; cancer is selected from one or more of colon cancer, breast cancer, thyroid cancer, alveolar rhabdomyosarcoma or hepatocellular carcinoma.

[0028] Additionally preferably, the complications of obesity are selected from one or more of hypertension; gallbladder disease; gastrointestinal disease; menstrual irregularities; degenerative arthritis; venous stasis ulcers; pulmonary hypoventilation syndrome; sleep apnea; snoring; coronary artery disease; arteriosclerotic disease; pseudotumor cerebri; accident proneness; increased surgical risk; osteoarthritis; high cholesterol; or an increased incidence of ovarian, cervical, uterine, breast, prostate or gallbladder malignancies.

[0029] In other aspects, the present invention also relates to the use and method of the above-mentioned compound, or a pharmaceutically acceptable salt, cocrystal, stereoisomer, solvate, prodrug or metabolite thereof for treating or preventing obesity or obesity complications. BRIEF DESCRIPTION OF THE DRAWINGS

[0030] Figure 1a shows the results of the inhibition of the CB1 receptor by the HTRF cAMP kit;

[0031] Figure 1b shows the results of the HTRF cAMP assay to determine the inhibitory effect of compounds Q1-P2 on CB1 receptors;

[0032] Figure 1c shows the results of the HTRF cAMP assay to determine the inhibitory effect of compound Q3-P2 on CB1 receptors;

[0033] Figure 1d shows the results of the inhibition of the control compound 2 on CB1 receptors determined by HTRF cAMP kit;

[0034] FIG2 a shows the results of the HTRF cAMP assay to determine the inhibitory effect of compound Q4-P1 on CB1 receptors;

[0035] FIG2 b shows the results of the HTRF cAMP assay to determine the inhibitory effect of compound Q5-P1 on CB1 receptors;

[0036] Figure 2c shows the results of the HTRF cAMP assay to determine the inhibitory effect of compound Q2-P2 on CB1 receptors;

[0037] Figure 2d shows the results of the HTRF cAMP assay to determine the inhibitory effect of compound Q6-P1 on CB1 receptors;

[0038] Figure 2e shows the results of the HTRF cAMP assay to determine the inhibitory effect of compound Q7-P1 on CB1 receptors;

[0039] Figure 2f shows the results of the HTRF cAMP assay to determine the inhibitory effect of compound Q8-P2 on CB1 receptors;

[0040] FIG3 a shows the results of the HTRF cAMP assay to determine the inhibitory effect of compound Q9-P1 on CB1 receptors;

[0041] FIG3 b shows the results of the HTRF cAMP assay to determine the inhibitory effect of compound Q11-P1 on CB1 receptors;

[0042] Figure 3c shows the results of the HTRF cAMP assay to determine the inhibitory effect of compound Q12-P1 on CB1 receptors;

[0043] Figure 3d shows the results of the HTRF cAMP assay to determine the inhibitory effect of compound Q13-P1 on CB1 receptors;

[0044] Figure 3e shows the results of the HTRF cAMP assay to determine the inhibitory effect of compound Q14-P2 on CB1 receptors;

[0045] FIG4 a shows the results of the HTRF cAMP assay to determine the inhibitory effect of compound Q10-P1 on CB1 receptors;

[0046] FIG4 b shows the results of the HTRF cAMP assay to determine the inhibitory effect of compound Q15-P2 on CB1 receptors;

[0047] Figure 4c shows the results of the inhibition of compound Q16-P1 on CB1 receptors determined by HTRF cAMP kit; and

[0048] FIG4 d shows the results of measuring the inhibitory effect of compound Q17-P2 on CB1 receptors using HTRF cAMP kit. DETAILED DESCRIPTION

[0049] Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which the claimed subject matter belongs.

[0050] "Pharmaceutically acceptable salt" or "pharmaceutically acceptable salt thereof" refers to a salt of the compound of the present invention that retains the biological effectiveness and properties of the free acid or free base, and the free acid is obtained by reacting with a non-toxic inorganic base or organic base, or the free base is obtained by reacting with a non-toxic inorganic acid or organic acid.

[0051] On the other hand, the term "pharmaceutically acceptable salt" herein refers to a salt that retains the desired biological activity of the subject compound and exhibits minimal undesirable toxicological effects, i.e., is suitable for use in contact with the tissues of a subject without excessive toxicity, irritation, allergic reaction, etc., within the scope of reasonable medical judgment, and is consistent with a reasonable benefit / risk ratio. These pharmaceutically acceptable salts can be prepared in situ during the final isolation and purification of the compound, or by reacting the free acid or free base form of the purified compound with a suitable base or acid, respectively. For example, Berge et al. describe pharmaceutically acceptable salts in detail in J. Pharmaceutical Sciences (1977) 66: 1-19. Pharmaceutically acceptable salts of the compounds provided herein include salts derived from suitable inorganic and organic acids and bases. Examples of pharmaceutically acceptable non-toxic acid addition salts are salts of amino groups formed with inorganic acids such as hydrochloric acid, hydrobromic acid, phosphoric acid, sulfuric acid and perchloric acid or organic acids such as acetic acid, oxalic acid, maleic acid, tartaric acid, citric acid, succinic acid or malonic acid, or by using other methods used in the art such as ion exchange. Other pharmaceutically acceptable salts include adipate, alginate, ascorbate, aspartate, benzenesulfonate, benzenesulfonate, benzoate, bisulfate, borate, butyrate, camphorate, camphorsulfonate, citrate, cyclopentanepropionate, digluconate, dodecylsulfate, ethanesulfonate, formate, fumarate, glucoheptonate, glycerophosphate, gluconate, hemisulfate, heptanoate, hexanoate, hydroiodide, 2-hydroxy-ethanesulfonate, lactobionic acid salt, lactate, laurate, lauryl sulfate, malate, maleate, malonate, methanesulfonate, 2-naphthalenesulfonate, naphthalene-m,n-disulfonate, nicotinate, nitrate, oleate, oxalate, palmitate, pamoate, pectinate, persulfate, 3-phenylpropionate, phosphate, picrate, pivalate, propionate, stearate, succinate, sulfate, tartrate, thiocyanate, p-toluenesulfonate, undecanoate, valerate, etc.

[0052] A "cocrystal" is a crystal formed by the active pharmaceutical ingredient (API) and cocrystal former (CCF) bound together by hydrogen bonds or other non-covalent bonds. Both the API and CCF are solid in their pure form at room temperature, and the components exist in a fixed stoichiometric ratio. A cocrystal is a multi-component crystal, encompassing both binary cocrystals formed between two neutral solids and multi-component cocrystals formed between a neutral solid and a salt or solvate.

[0053] "Stereoisomers" refer to isomers resulting from different spatial arrangements of atoms in a molecule, including cis-trans isomers, enantiomers, and conformational isomers.

[0054] "Solvate" or "solvate" refers to a solvent addition form containing either a stoichiometric or non-stoichiometric amount of solvent. Some compounds tend to trap fixed molar ratios of solvent molecules in their crystalline solid state, thereby forming solvates. If the solvent is water, the solvate formed is a hydrate; if the solvent is an alcohol, the solvate formed is an alcoholate. Hydrates are formed by the association of one or more water molecules with one molecule of the substance, where the water remains in its molecular form as HO.

[0055] "Prodrugs" refer to compounds of the present invention that can be converted into biologically active compounds through in vivo metabolism. Prodrugs of the present invention are prepared by modifying amino or carboxyl groups in compounds of the present invention. These modifications can be removed by conventional manipulation or in vivo to yield the parent compound. When the prodrugs of the present invention are administered to a mammalian subject, the prodrugs are cleaved to form free amino or carboxyl groups. Non-limiting examples of prodrugs include esters, carbonates, half-esters, phosphates, nitroesters, sulfates, sulfoxides, amides, carbamates, nitrogen-containing compounds, phosphamides, glycosides, ethers, acetals, and ketals.

[0056] A "metabolite" of a compound disclosed herein is a derivative of the compound formed when the compound is metabolized. The term "active metabolite" refers to a biologically active derivative of a compound formed when the compound is metabolized. The term "metabolized," as used herein, refers to the sum of processes by which a particular substance is altered by an organism (including, but not limited to, hydrolysis reactions and reactions catalyzed by enzymes, such as oxidation reactions). Thus, an enzyme can produce a specific structural transformation into a compound. For example, cytochrome P450 catalyzes various oxidation and reduction reactions, while diphosphoglucosyltransferase catalyzes the conversion of activated glucuronic acid molecules to aromatic alcohols, aliphatic alcohols, carboxylic acids, amines, and free sulfhydryl groups. Further information on metabolism can be obtained from "The Pharmacological Basis of Therapeutics," 9th edition, McGraw-Hill (1996). Metabolites of the compounds disclosed herein can be identified by administering the compound to a host and analyzing tissue samples from the host, or by incubating the compound with hepatocytes in vitro and analyzing the resulting compounds. Both methods are known in the art. In some embodiments, metabolites of the compound are formed by an oxidation process and correspond to the corresponding hydroxyl-containing compound. In some embodiments, the compound is metabolized to a pharmaceutically active metabolite.

[0057] "Pharmaceutical composition" refers to a mixture of one or more compounds of the present invention, their pharmaceutically acceptable salts or prodrugs and other chemical components, wherein "other chemical components" refers to pharmaceutically acceptable carriers, excipients and / or one or more other therapeutic agents.

[0058] "Carrier" refers to a material that does not cause significant irritation to an organism and does not abrogate the biological activity and properties of the administered compound.

[0059] "Excipient" refers to an inert substance added to a pharmaceutical composition to facilitate administration of a compound. Non-limiting examples include calcium carbonate, calcium phosphate, sugars, starches, cellulose derivatives (including microcrystalline cellulose), gelatin, vegetable oils, polyethylene glycols, diluents, granulating agents, lubricants, binders, and disintegrants.

[0060] An "effective amount" refers to an amount of a drug or pharmaceutical formulation that will elicit the biological or medical response of a tissue, system, animal, or human being, for example, that is being studied by a researcher or physician. Furthermore, the term "therapeutically effective amount" refers to any amount that results in treatment, cure, prevention, or alleviation of a disease, disorder, or side effect, or a reduction in the rate of progression of a disease or disorder, compared to a corresponding subject that has not received that amount. Also included within the scope of the term is an amount effective to enhance normal physiological function.

[0061] As used herein, the term "treating" refers to alleviating at least one symptom of a disease, disorder, or condition. The term includes administering and / or applying one or more compounds described herein to a subject to provide management or treatment of a condition. "Treatment" for the purposes of this disclosure may, but does not necessarily, provide a cure; rather, it is meant that "treatment" can be a form of management of a condition. When the compounds described herein are used to treat harmful proliferating cells (including cancer), "treatment" includes partial or complete destruction of the harmful proliferating cells with minimal damage to normal cells. The desired treatment mechanism for harmful rapidly proliferating cells (including cancer cells) at the cellular level is apoptosis.

[0062] As used herein, the term "prevention" includes both preventing or slowing the onset of clinically significant disease development or preventing or slowing the onset of a preclinically significant disease stage in an at-risk individual. This includes prophylactic treatment of individuals at risk of developing disease.

[0063] The term "subject" or "patient" includes organisms that can suffer from a disorder or a disorder associated with reduced or insufficient programmed cell death (apoptosis) or that can otherwise benefit from the administration of the compounds of the invention, such as humans and non-human animals. Preferred humans include human patients suffering from or prone to suffering from a disorder or related condition as described herein. The term "non-human animal" includes vertebrates, such as mammals, such as non-human primates, sheep, cattle, dogs, cats, and rodents such as mice, as well as non-mammals, such as chickens, amphibians, reptiles, etc.

[0064] The GI used in this paper 50 It refers to the drug concentration required to inhibit 50% of cell growth, that is, the drug concentration when the growth of 50% of cells (such as cancer cells) is inhibited or controlled.

[0065] IC used in this article 50 It refers to the amount, concentration, or dose of a particular test compound that achieves 50% inhibition of the maximal effect in the assay in which the effect is measured.

[0066] The EC used in this paper 50 It refers to the dose, concentration or amount of a test compound that elicits a dose-dependent response that elicits 50% of the maximal expression of a specific response induced, stimulated or potentiated by the particular test compound.

[0067] Unless otherwise indicated, the present invention employs conventional methods such as mass spectrometry, NMR, HPLC, protein chemistry, biochemistry, recombinant DNA technology, and pharmacology within the skill of the art. Unless specific definitions are provided, the nomenclature and laboratory procedures and techniques associated with analytical chemistry, synthetic organic chemistry, and medical and pharmaceutical chemistry described herein are those known to those skilled in the art. In general, the aforementioned techniques and steps can be implemented by conventional methods well known in the art and described in various general and more specific literature, which are cited and discussed in this specification.

[0068] Active compound

[0069] The present disclosure relates to a CB1 inhibitor, specifically to a compound, or a pharmaceutically acceptable salt, cocrystal, stereoisomer, solvate, prodrug or metabolite thereof, wherein the compound is selected from:

[0070] The present disclosure also relates to compounds of formula (I), or pharmaceutically acceptable salts, cocrystals, stereoisomers, solvates, prodrugs or metabolites thereof:

[0071] in,

[0072] Ring A represents a five-membered or six-membered monocyclic heteroaryl group containing 1 or 2 heteroatoms independently selected from N and S, preferably selected from thienyl, pyridinyl and thiazolyl, more preferably selected from thien-2-yl, pyridin-2-yl and thiazol-5-yl;

[0073] R1 is C 1-6 Halogenated alkyl, preferably C 1-3 Haloalkyl, more preferably C 1-3 Fluoroalkyl, most preferably trifluoromethyl;

[0074] m is 0, 1 or 2, more preferably 1 or 2, particularly preferably 1;

[0075] R2 are each independently selected from halogen, cyano, and C 1-6 Halogenated alkyl groups are preferably each independently selected from fluorine, chlorine, cyano, and C 1-3 Halogenated alkyl, particularly preferably C 1-3 Fluoroalkyl, more preferably trifluoromethyl;

[0076] R 3a 、R 3b 、R 4a and R 4b are each independently selected from hydrogen and C 1-6 Alkyl groups are preferably each independently selected from hydrogen and C 1-3Alkyl, more preferably each independently selected from hydrogen and methyl, most preferably R 3a and R 3b At least one of them is hydrogen, and R 4a and R 4b Each is hydrogen.

[0077] The present disclosure also relates to compounds of formula (II), or pharmaceutically acceptable salts, cocrystals, stereoisomers, solvates, prodrugs or metabolites thereof:

[0078] in,

[0079] Ring B represents a phenyl group or a five-membered or six-membered monocyclic heteroaryl group containing 1 or 2 heteroatoms independently selected from N and S, preferably selected from phenyl, thienyl, and pyridinyl, more preferably selected from phenyl, thien-2-yl, and pyridin-2-yl;

[0080] Ring C represents phenyl or thienyl, more preferably selected from phenyl and thien-2-yl;

[0081] R1 is C 1-6 Halogenated alkyl, preferably C 1-3 Haloalkyl, more preferably C 1-3 Fluoroalkyl, most preferably trifluoromethyl;

[0082] m is 0, 1 or 2, more preferably 1 or 2, particularly preferably 1;

[0083] R2 are each independently selected from halogen, cyano, and C 1-6 Halogenated alkyl groups are preferably each independently selected from fluorine, chlorine, cyano, and C 1-3 Halogenated alkyl, particularly preferably C 1-3 Fluoroalkyl, more preferably trifluoromethyl;

[0084] R 3a 、R 3b 、R 4a and R 4b are each independently selected from hydrogen and C 1-6 Alkyl groups are preferably each independently selected from hydrogen and C 1-3 Alkyl, more preferably each independently selected from hydrogen and methyl, most preferably R 3a and R 3b At least one of them is hydrogen, and R 4a and R 4b Each is hydrogen.

[0085] Also described herein are pharmaceutically acceptable salts, cocrystals, stereoisomers, solvates, prodrugs, or metabolites of this compound.

[0086] In particular, the compounds described herein can be made and / or used as pharmaceutically acceptable salts. Types of pharmaceutically acceptable salts include, but are not limited to: (1) acid addition salts, formed by reacting the free base form of the compound with a pharmaceutically acceptable inorganic acid, such as hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid, phosphoric acid, metaphosphoric acid, or the like; or with an organic acid, such as acetic acid, propionic acid, hexanoic acid, cyclopentanepropionic acid, glycolic acid, pyruvic acid, lactic acid, malonic acid, malic acid, citric acid, succinic acid, maleic acid, tartaric acid, fumaric acid, trifluoroacetic acid, benzoic acid, 3-(4-hydroxybenzoyl)benzoic acid, cinnamic acid, mandelic acid, methanesulfonic acid, ethanesulfonic acid, 1,2-ethanedisulfonic acid, 2-hydroxyethanesulfonic acid, benzenesulfonic acid, toluenesulfonic acid, 4-methylbicyclo-[2.2.2]oct-2-ene-1- -formic acid, 2-naphthalenesulfonic acid, tert-butylacetic acid, glucoheptonic acid, 4,4'-methylenebis-(3-hydroxy-2-ene-1-carboxylic acid), 3-phenylpropionic acid, trimethylacetic acid, dodecylsulfuric acid, gluconic acid, glutamic acid, salicylic acid, hydroxynaphthoic acid, stearic acid, muconic acid, etc.; (2) base addition salts, which are formed when the acidic protons in the parent compound are replaced by metal ions, such as alkali metal ions (such as lithium, sodium, potassium), alkaline earth metal ions (such as magnesium or calcium) or aluminum ions; or coordinated with organic bases or inorganic bases, acceptable organic bases include ethanolamine, diethanolamine, triethanolamine, trimethylamine, N-methylglucamine, etc.; acceptable inorganic bases include aluminum hydroxide, calcium hydroxide, potassium hydroxide, sodium carbonate, sodium hydroxide, etc.

[0087] The corresponding counter ion of the pharmaceutically acceptable salt can be analyzed and identified using various methods including, but not limited to, ion exchange chromatography, ion chromatography, capillary electrophoresis, inductively coupled plasma, atomic absorption spectroscopy, mass spectrometry, or any combination thereof.

[0088] The salt is recovered using at least one of the following techniques: filtration, precipitation with a non-solvent followed by filtration, evaporation of the solvent, or, in the case of aqueous solutions, lyophilization.

[0089] Screening and characterizing pharmaceutically acceptable salts, polymorphs and / or solvates can be accomplished using a variety of techniques including, but not limited to, thermal analysis, X-ray diffraction, spectroscopy, microscopy, elemental analysis. The various spectroscopic techniques used include, but are not limited to, Raman, FTIR, UVIS, and NMR (liquid and solid state). Various microscopic techniques include, but are not limited to, IR microscopy and Raman microscopy.

[0090] Drug uses

[0091] The CB1 inhibitors of the present disclosure can be used to treat or prevent obesity or complications of obesity.

[0092] In a preferred embodiment, the complications of obesity are selected from one or more of diabetes, dyslipidemia, metabolic syndrome, dementia, inflammatory disease, cardiovascular disease, liver disease, or cancer.

[0093] Further preferably, diabetes is selected from one or more of type 1 diabetes, type 2 diabetes, impaired glucose tolerance or insulin resistance; dyslipidemia is selected from one or more of poor blood lipid levels, low levels of high-density lipoprotein, high levels of low-density lipoprotein or high levels of triglycerides; inflammatory diseases are selected from one or more of osteoarthritis, rheumatoid arthritis, inflammatory bowel disease or obesity-related inflammation; cardiovascular diseases are selected from one or more of atherosclerosis, hypertension, stroke or heart attack; liver disease is selected from one or more of liver inflammation, liver fibrosis, non-alcoholic steatohepatitis, fatty liver, liver enlargement, alcoholic liver disease, jaundice, cirrhosis or hepatitis; cancer is selected from one or more of colon cancer, breast cancer, thyroid cancer, alveolar rhabdomyosarcoma or hepatocellular carcinoma.

[0094] Additionally preferably, the complications of obesity are selected from one or more of hypertension; gallbladder disease; gastrointestinal disease; menstrual irregularities; degenerative arthritis; venous stasis ulcers; pulmonary hypoventilation syndrome; sleep apnea; snoring; coronary artery disease; arteriosclerotic disease; pseudotumor cerebri; accident proneness; increased surgical risk; osteoarthritis; high cholesterol; or an increased incidence of ovarian, cervical, uterine, breast, prostate or gallbladder malignancies.

[0095] In an embodiment of the present invention, the medicine comprising the compounds of this invention can be administered to the patient by at least one of injection, oral administration, inhalation, rectal administration and transdermal administration. When the patient is treated according to the present invention, the amount of a given drug depends on many factors, such as specific dosage regimen, disease or disease type and severity thereof, the uniqueness (such as body weight) of the patient or host for treatment, but, according to specific surrounding circumstances, including, for example, the specific drug, route of administration, disease for treatment and the patient or host for treatment, the dosage can be conventionally determined by methods known in the art. Usually, for the dosage used in adult treatment, the dosage is typically 0.02-5000mg / days, such as the scope of about 1-1500mg / days. The desired dose can be conveniently expressed as a dose or simultaneously administered (or in a short time) or at appropriate intervals, such as two, three, four or more doses per day. It will be appreciated by those skilled in the art that, although the above-mentioned dosage range has been given, specific effective amount can be appropriately adjusted according to the patient's situation and in conjunction with a physician's diagnosis.

[0096] In some embodiments of the methods or uses disclosed herein, a compound as described herein, any one of the compounds, is administered to a subject at a dosage (e.g., a therapeutically effective dose) of about 2 mg, 1-3 mg, 1-5 mg, 1-10 mg, 0.5-20 mg, or 0.1-50 mg. In some embodiments, the dosage (e.g., a therapeutically effective dose) is about 2 mg, 1-3 mg, 1-5 mg, 1-10 mg, 0.5-20 mg, 0.1-50 mg, 0.1-75 mg, 0.5-75 mg, 1-75 mg, 0.1-100 mg, 0.5-100 mg, or 1-100 mg. In some embodiments, the dosage is about 1-10 mg. In some embodiments, the dosage is about 1-50 mg. In some embodiments, the dosage is about 1-100 mg.

[0097] Pharmaceutical composition

[0098] Another aspect of the present invention relates to a pharmaceutical composition comprising (1) a compound as described above, or a pharmaceutically acceptable salt, cocrystal, stereoisomer, solvate, prodrug or metabolite thereof; (2) optionally one or more other active ingredients; and (3) a pharmaceutically acceptable carrier and / or excipient.

[0099] Preparation of compounds

[0100] The compounds of the present invention can be synthesized using standard synthetic techniques known to those skilled in the art or using methods known in the art in combination with the methods described herein. In addition, the solvents, temperatures, and other reaction conditions given herein can be varied according to the skill in the art. As further guidance, the following synthetic methods can also be utilized.

[0101] The reactions can be used sequentially to provide the compounds described herein; or they can be used to synthesize fragments that are subsequently added by methods described herein and / or known in the art.

[0102] The starting materials for synthesizing the compounds described herein can be synthesized or obtained from commercial sources. The compounds described herein and other related compounds having different substituents can be synthesized using techniques and raw materials known to those skilled in the art. General methods for preparing the compounds disclosed herein can be derived from reactions known in the art, and the reactions can be modified by reagents and conditions deemed appropriate by those skilled in the art to introduce various moieties into the molecules provided herein.

[0103] If desired, the reaction products can be isolated and purified using conventional techniques, including but not limited to filtration, distillation, crystallization, chromatography, etc. These products can be characterized using conventional methods, including physical constants and spectral data.

[0104] Non-limiting examples of synthetic schemes for preparing compounds of the present invention are described below.

[0105] Preparation Example 1. Exemplary Synthesis of 3-(4-chlorophenyl)-4-phenyl-N-(2-sulfamoylethyl)-N′-((4-(cyclopropyl)phenyl)sulfonyl)-4,5-dihydro-1H-pyrazole-1-carboximidamide (Compound Q1)

[0106] A solution of 4-cyclopropylbenzenesulfonyl chloride in THF was bubbled with nitrogen for 20 minutes, and the reaction product was then passed through a pad of silica gel and concentrated to afford 4-cyclopropylbenzenesulfonamide.

[0107] To a solution of 4-cyclopropylbenzenesulfonamide in acetonitrile was added triethylamine (2.5 equivalents) and methyl chloroformate (1.5 equivalents) at 0°C, and the reaction system was slowly warmed to room temperature and stirred overnight. After completion of the reaction, water was added to quench the reaction, and the mixture was concentrated under reduced pressure to yield methyl ((4-cyclopropylphenyl)sulfonyl)carbamate.

[0108] An equivalent amount of 3-(4-chlorophenyl)-4-phenyl-4,5-dihydro-1H-pyrazole was added to a toluene solution of methyl ((4-cyclopropylphenyl)sulfonyl)carbamate, and the mixture was heated to 110°C and stirred overnight. After completion of the reaction, the mixture was cooled to room temperature and concentrated under reduced pressure to obtain N-(3-(4-chlorophenyl)-4-phenyl-4,5-dihydro-pyrazole-1-carbonyl)-4-cyclopropyl-benzenesulfonamide.

[0109] To a toluene solution of N-(3-(4-chlorophenyl)-4-phenyl-4,5-dihydro-pyrazol-1-carbonyl)-4-cyclopropyl-benzenesulfonamide, N,N-diisopropylethylamine (1.5 equivalents) and phosphorus oxychloride (1.3 equivalents) were added, and the mixture was stirred at 85°C for 15 hours. After the reaction was completed, the mixture was cooled to room temperature and concentrated under reduced pressure to obtain N-(chloro-(3-(4-chloro-phenyl)-4-phenyl-4,5-dihydro-pyrazol-1-yl)-methylene)-4-cyclopropyl-benzenesulfonamide.

[0110] To a dichloromethane solution of N-(chloro-(3-(4-chloro-phenyl)-4-phenyl-4,5-dihydro-pyrazol-1-yl)-methylene)-4-cyclopropyl-benzenesulfonamide was added triethylamine (1 equivalent) and 2-aminoethanesulfonamide (1.5 equivalents), and the mixture was stirred at room temperature overnight. After completion of the reaction, the mixture was concentrated under reduced pressure to yield 3-(4-chlorophenyl)-4-phenyl-N-(2-sulfamoylethyl)-N'-((4-(cyclopropyl)phenyl)sulfonyl)-4,5-dihydro-1H-pyrazole-1-carboximidamide (Compound Q1).

[0111] Preparation Example 2-17

[0112] Compounds Q2-Q17 were synthesized similarly to compound Q1.

[0113] Preparation Example 18. Synthesis of (Z)-3-(4-chlorophenyl)-N'-((4-cyclopropylphenyl)sulfonyl)-4-phenyl-N-(2-sulfamoylethyl)-4,5-dihydro-1H-pyrazole-1-carboximide (S, R enantiomers of Compound Q1)

[0114] In addition to the preparation method shown in Preparation Example 1, the enantiomer of Compound Q1 can be synthesized by the following method.

[0115] At 0 ° C, compound 1 (100 mg, 424 μmol) and Et3N (107 mg, 1.06 mmol, 147 μL) in DCM (2.00 mL) was added compound 1a (79.6 mg, 508 μmol, 63.8 μL). The mixture was stirred at 0 ° C for 4 hours. TLC (dichloromethane: methanol = 10: 1, Rf = 0.40) showed that compound 1 was completely consumed. The reaction mixture was diluted with DCM (5.00 mL) and washed with H2O (5.00 mL × 2), brine (5.00 mL), and dried over Na2SO4. The organic layer was concentrated under reduced pressure at 35 ° C. The residue was purified by flash silica gel chromatography ( 4g The product was purified by silica gel flash column, 0-10% dichloromethane / methanol eluent @ 30 mL / min) to afford yellow solid compound 2 (50.0 mg, 84.2 μmol, 19.9% ​​yield, 60.0% purity).

[0116] 1 H NMR: (400MHz, CDCl3) δ7.84-7.91(m,2H),7.65(br d,J=8.4Hz,1H),7.54(br d,J=8.4Hz,2H),7.07-7.16(m,1H),6.98(br d,J=8.0Hz,2H),6.83(br d,J=7.6Hz,1H).

[0117] Compound 2 (1.50 g, 4.21 mmol) and compound 2a (1.19 g, 4.63 mmol) were degassed in toluene (20.0 mL) and purged with N2 three times, and then the mixture was stirred at 110 ° C under N2 atmosphere for 1 hour. TLC (petroleum ether: ethyl acetate = 3: 1, product: Rf = 0.30) showed that compound 2 was completely consumed. The reaction mixture was concentrated under reduced pressure to remove the solvent. The residue was purified by flash silica gel chromatography ( 12g The product was purified by silica gel flash column with a gradient of 0-50% ethyl acetate / petroleum ether at 50 mL / min as eluent to afford compound 3 as a yellow solid (1.10 g, 1.91 mmol, 45.3% yield, 90.0% purity).

[0118] 1 H NMR: (400MHz, CDCl3) δ8.76(br s,1H),8.03(br d,J=8.4Hz,2H),7.69(br d,J=8.4Hz,2H),7.53(br d,J=8.4Hz,2H),7.27-7.36(m,4H),7.11(br d, J=7.2Hz, 2H), 4.71 (br dd, J=11.6, 5.4Hz, 1H), 4.31 (t, J=11.6Hz, 1H), 3.90 (dd, J=11.4, 5.6Hz, 1H).

[0119] Compound 3 (500 mg, 964 μmol), Pd (dppf) Cl (70.5 mg, 96.4 μmol), Na CO (511 mg, 4.82 mmol) and compound 3a (82.8 mg, 964 μmol) in toluene (6.00 mL) and H O (2.00 mL) were degassed and purged with N 3 times, and the mixture was then stirred at 100 ° C under N atmosphere for 3 hours. TLC (petroleum ether: ethyl acetate = 3: 1, product: Rf = 0.30) showed that compound 3 was completely consumed. The reaction mixture was diluted with H O (5.00 mL) and extracted with ethyl acetate (10.0 mL × 3). The combined organic layers were washed with brine (10.0 mL × 2), dried over Na SO , filtered and concentrated under reduced pressure to give compound 4 (493 mg, 822 μmol, 85.3% yield, 80.0% purity) as a brown solid.

[0120] 1H NMR: (400MHz, CDCl3) δ8.70(br s,1H),8.02(d,J=8.4Hz,1H),7.49-7.56(m,3H),7.27-7.36(m,4H),7.20(br d,J=8.4Hz,3H),7.10(br d,J=6.8Hz,3H),4.69(dd,J=11.6,5.4Hz,1H),4.29(br t,J=11.6Hz,1H),3.89(br dd,J=11.6,5.5Hz,1H),1.87-2.01(m,1H),1.04-1.14(m,2H),0.75-0.83(m,2H).

[0121] Toluene (5.00mL) solution of POCl (256mg, 1.67mmol, 155μL) was added DIEA (215mg, 1.67mmol, 290μL) and compound 4 (400mg, 833μmol). The mixture was stirred at 100°C under N atmosphere for 2 hours. LCMS showed that compound 4 was completely consumed and obtained two main peaks (Rt=3.082min and Rt=3.124min). The reaction mixture was quenched with H o (5.00mL) and extracted with ethyl acetate (10.0mL×3). The combined organic layers were washed with brine (10.0mL×2), dried over Na sO , filtered and concentrated under reduced pressure to give compound 5 (440mg, 706μmol, 84.7% yield, 80.0% purity) as a brown solid.

[0122] LCMS: m / z=498.0 (M+H) + ,Rt=3.124min.

[0123] LCMS: m / z=498.0 (M+H) + ,Rt=3.082min.

[0124] To a solution of compound 5 (380 mg, 762 μmol) in DCM (5.00 mL) was added Et3N (386 mg, 3.81 mmol, 531 μL) and compound 5a (184 mg, 1.14 mmol). The mixture was stirred at 25 ° C for 2 hours. TLC (petroleum ether (containing 0.3% NH4OH): ethyl acetate = 1: 1, Rf = 0.25) showed that compound 5 was completely consumed and two new spots were formed. The reaction mixture was concentrated under reduced pressure to obtain a residue. The residue was purified by flash silica gel chromatography ( 12g The product was purified by silica gel flash column, 0-50% ethyl acetate / petroleum ether gradient elution @ 50 mL / min), and then purified by SFC (column: (s,s) WHELK-O1 (250 mm*30 mm, 10 um); mobile phase: [CO2-ACN / i-PrOH (0.1% NH3H2O)]; B%: 45%, isocratic elution mode) to give compound Q1-P1 (55.0 mg, 99.37% purity) as a white solid and compound Q1-P2 (55.0 mg, 97.82% purity) as a white solid.

[0125] Compound Q1-P1:

[0126] LCMS: m / z=586.1 (M+H) + ,Rt=2.634min.

[0127] 1 H NMR: (400MHz, CDCl3) δ7.77(d,J=8.4Hz,2H),7.52(d,J=8.6Hz,2H),7.47(br t,J=6.0Hz,1H),7.27-7.36(m,3H),7.24(s,1H),7.05-7.15(m,4H),4.95(s,2H),4.62-4.73(m,1H),4.48-4.59(m,1H),4.18( q,J=6.3Hz,2H),4.10(dd,J=11.6,5.4Hz,1H),3.50(t,J=6.1Hz,2H),1.85-2.02(m,1H),0.98-1.07(m,2H),0.65-0.76(m,2H).

[0128] Compound Q1-P2:

[0129] LCMS: m / z=586.1 (M+H) + ,Rt=2.633min.

[0130] 1H NMR: (400MHz, CDCl3) δ7.77(d,J=8.4Hz,2H),7.52(d,J=8.6Hz,2H),7.47(br t,J=6.0Hz,1H),7.27-7.36(m,3H),7.24(s,1H),7.05-7.15(m,4H),4.95(s,2H),4.62-4.73(m,1H),4.48-4.59(m,1H),4.18( q,J=6.3Hz,2H),4.10(dd,J=11.6,5.4Hz,1H),3.50(t,J=6.1Hz,2H),1.85-2.02(m,1H),0.98-1.07(m,2H),0.65-0.76(m,2H).

[0131] Preparation Example 19. Synthesis of (Z)-3-(5-chlorothiophen-2-yl)-4-phenyl-N-(2-sulfamoylethyl)-N'-((4-(trifluoromethyl)phenyl)sulfonyl)-4,5-dihydro-1H-pyrazole-1-carboximide (S and R enantiomers of Compound Q3)

[0132] At 0 ° C, AlCl (5.94g, 44.5mmol, 2.44mL) was added to a mixture of compound 1 (5.30g, 34.2mmol, 4.57mL) and compound 1a (4.07g, 34.2mmol, 3.16mL) in DCM (50.0mL). The reaction mixture was stirred at 25 ° C under N2 atmosphere for 1 hour. TLC (petroleum ether: ethyl acetate = 3: 1, Rf = 0.50) showed that compound 1 was completely consumed. The reaction mixture was quenched with 1M HCl (50.0mL) and extracted with DCM (100mL × 3). The combined organic layer was washed with brine (100mL × 2), dried over Na2SO4, filtered and concentrated under reduced pressure to give a residue. The residue was purified by flash silica gel chromatography ( 80g The product was purified by silica gel flash column with a gradient of 0-30% ethyl acetate / petroleum ether at 60 mL / min as the eluent to afford compound 2 (5.1 g, 20.4 mmol, 59.7% yield, 95% purity) as a yellow solid.

[0133] 1 H NMR: (400MHz, CDCl3) δ7.51 (d, J = 4.1 Hz, 1H), 7.27-7.33 (m, 2H), 7.21-7.27 (m, 3H), 6.91 (d, J = 4.1 Hz, 1H), 4.09 (s, 2H).

[0134] To a solution of compound 2 (2.00 g, 8.45 mmol) and formaldehyde (3.43 g, 42.2 mmol, 3.15 mL) in MeOH (20.0 mL) was added piperidine (719 mg, 8.45 mol, 834 μL) and AcOH (1.01 g, 16.9 mmol, 967 μL). The mixture was stirred at 70 ° C for 2 hours. LCMS (Rt = 1.560 minutes) showed that compound 2 was completely consumed. The reaction mixture was filtered and concentrated under reduced pressure to give compound 3 (2.10 g, crude product) as a yellow oil.

[0135] LCMS: m / z=249.2 (M+H) + ,Rt=1.561min.

[0136] Compound 3 (2.10g, 8.44mmol) and NH2NH2·H2O (4.27g, 72.5mmol, 4.14mL) in EtOH (21.0 milliliters) solution was stirred at 80 ℃ for 3 hours.LCMS (Rt=1.216 minutes) showed that compound 3 was completely consumed. The reaction mixture was concentrated under reduced pressure to obtain a residue. Then EtOH (20mL) was added to the residue at 0 ℃, and a yellow solid precipitate was observed. It was filtered through a sand core funnel and washed with EtOH (5mL×3). The yellow filter cake was concentrated under reduced pressure to obtain INT_4 (970mg, 3.51mmol, 41.5% yield, 95% purity) as a white solid.

[0137] LCMS: m / z=263.2 (M+H) + ,Rt=1.216min.

[0138] 1 H NMR: (400MHz, CDCl3) δ7.17-7.28(m,5H),6.56(d,J=4.0Hz,1H),6.39(d,J=4.0Hz,1H),5.59(br s, 1H), 4.34 (dd, J = 10.6, 7.1Hz, 1H), 3.92 (t, J = 10.1Hz, 1H), 3.45 (dd, J = 9.4, 7.2Hz, 1H).

[0139] To a solution of compound 5 (2.00 g, 8.88 mmol) and TEA (2.25 g, 22.2 mmol, 3.09 mL) in DCM (20.0 mL) was added compound 5a (1.67 g, 10.6 mmol, 1.34 ml) at 0°C. The mixture was stirred at 0°C for 4 hours. LCMS (Rt = 1.335 minutes) showed that compound 5 was completely consumed. H2O (30.0 mL) was added at 0°C to quench the reaction mixture, which was then extracted with DCM (20.0 mL × 3). The combined organic layers were washed with brine (30.0 mL), dried over Na2SO4, filtered and concentrated under reduced pressure to give compound 6 (3.07 g, 8.00 mmol, 90.1% yield, 90% purity) as a white solid.

[0140] LCMS: m / z=346.1 (M+H) + ,Rt=1.335min.

[0141] 1 H NMR: (400MHz, CDCl3) δ8.13 (d, J = 8.1 Hz, 2H), 7.68 (d, J = 8.1 Hz, 2H), 7.24-7.32 (m, 3H), 7.08-7.15 (m, 1H), 6.98 (d, J = 8.3 Hz, 2H).

[0142] To a solution of compound 6 (600 mg, 1.74 mmol) in toluene (6.00 ml) was added INT-4 (456 mg, 1.74 mmol). The mixture was stirred at 110° C. for 2 hours. LCMS (Rt=1.694 minutes) showed that compound 6 was completely consumed. The reaction mixture was filtered and concentrated under reduced pressure to give compound 7 (890 mg, 1.73 mmol, 99.6% yield) as a yellow oil.

[0143] LCMS: m / z=514.1 (M+H) + ,Rt=1.694min.

[0144] To a toluene (9.00 milliliters) solution of compound 7 (890 mg, 1.73 mmol) was added POCl (531 mg, 3.46 mmol, 322 μL) and DIEA (447 mg, 3.46 mm mol, 603 μL). The mixture was stirred at 110° C. for 2 hours. LCMS (Rt=1.818 minutes) showed that compound 7 was completely consumed. H was added at 20° C. O (50.0 mL) to quench the reaction mixture, which was then extracted with ethyl acetate (50.0 mL×3). The combined organic layers were washed with brine (50.0 mL), dried over Na SO, filtered, and concentrated under reduced pressure to afford compound 8 (921 mg, 1.73 mmol, 99.9% yield) as a yellow solid.

[0145] LCMS: / z=532.1(M+H) + ,Rt=1.818min.

[0146] To a solution of compound 8 (600 mg, 1.13 mmol) in DCM (6.00 mL) was added TEA (570 mg, 5.64 mmol, 784.3 μL) and compound 8a (271.53 mg, 1.69 mmol). The mixture was stirred at 25 ° C for 2 hours. LCMS (Rt = 1.568 minutes) showed that compound 8 was completely consumed, and a major peak with the desired mass was detected (TLC petroleum ether: ethyl acetate = 1:1, R f =0.1). The reaction mixture was filtered and concentrated under reduced pressure to give a residue. The residue was first purified by flash silica gel chromatography ( 20g The residue was purified by silica gel flash column (0-50% petroleum ether:ethyl acetate gradient elution @ 40 mL / min). The residue was then purified by SFC (column: DAICEL CHIRALCEL OD (250 mm × 30 mm, 10 μm); mobile phase: [CO₂-MeOH (0.1% NH₃H₂O)]; B%: 30%, isocratic elution mode) to obtain compound Q3-P1 (44.21 mg, 96.46% purity) as a white solid and compound Q3-P2 (36.19 mg, 99.75% purity) as a white solid.

[0147] Compound Q3-P1:

[0148] LCMS: m / z=620.3 (M+H) + ,Rt=1.570min.

[0149] 1H NMR: (400MHz, CDCl3) δ8.03(d,J=8.3Hz,2H),7.68(d,J=8.4Hz,2H),7.49(br t,J=6.0Hz,1H),7.28-7.37(m,3H),7.13-7.20(m,2H),6.69(s,2H),5.13(s,2H),4.51- 4.64(m,2H), 4.14(q,J=6.0Hz,2H), 4.05(dd,J=10.4,5.2Hz,1H), 3.51(t,J=6.0Hz,2H).

[0150] Compound Q 3-P2:

[0151] LCMS: m / z=620.2 (M+H) + ,Rt=1.572min.

[0152] 1 H NMR: (400MHz, CDCl3) δ8.03(d,J=8.3Hz,2H),7.69(d,J=8.4Hz,2H),7.47(br t,J=6.0Hz,1H),7.28-7.38(m,3H),7.13-7.20(m,2H),6.69(s,2H),5.06(s,2H),4.51- 4.65(m,2H), 4.14(q,J=6.0Hz,2H), 4.06(dd,J=10.4,5.2Hz,1H), 3.51(t,J=6.0Hz,2H).

[0153] Preparation Example 20: Synthesis of (Z)-3-(4-chlorophenyl)-4-phenyl-N-(2-sulfamoylethyl)-N'-((5-(trifluoromethyl)thiophen-2-yl)sulfonyl)-4,5-dihydro-1H-pyrazole-1-carboximide (S, R enantiomers of compound Q4)

[0154] A mixture of compound 1 (2.00 g, 6.71 mmol), CuI (127 mg, 670 μmol) and compound 1b (1.55 g, 8.05 mmol, 1.02 mL) dissolved in DMF (20.0 mL) was degassed and purged with N2 three times, and then the mixture was stirred at 100°C under N2 atmosphere for 12 hours. TLC (petroleum ether:ethyl acetate = 3:1, R f =0.3) indicated that compound 1 was completely consumed and a new spot was formed. The reaction mixture was filtered and concentrated under reduced pressure to obtain a residue. The residue was purified by flash silica gel chromatography ( 40g The product was purified by silica gel flash column (0-50% ethyl acetate / petroleum ether gradient @ 50 mL / min) to afford compound 2 (1.70 g, 5.92 mmol, crude product) as a yellow oil.

[0155] 1 H NMR: (400MHz, CDCl3-d) δ7.35 (d, J=4.0Hz, 1H), 7.01 (d, J=4.0Hz, 1H), 1.31 (s, 9H).

[0156] A mixture of compound 2 (500 mg, 1.74 mmol) and TFA (3.07 g, 26.9 mmol, 2.00 mL) dissolved in DCM (0.60 mL) was degassed and purged with N 2 three times, and the mixture was then stirred at 25 ° C under an N 2 atmosphere for 12 hours. LCMS (Rt=1.379 min) showed that compound 2 was completely consumed, and a main peak with the desired mass number was detected. The reaction mixture was filtered and concentrated under reduced pressure to give INT_2 (400 mg, 1.73 mmol, crude product) as a yellow oil.

[0157] LCMS: m / z = 232.0 (M+H) + , Rt=1.379min.

[0158] Compound INT_2 (400 mg, 1.73 mmol) and TEA (437 mg, 4.32 mmol, 601 μL) were dissolved in DCM (4.00 mL) at 0°C. Compound 2a (325 mg, 2.08 mmol, 260 μL) was then added dropwise at 0°C. The mixture was stirred at 0°C for 4 hours. LCMS (Rt = 1.785 min) showed that compound 2 was completely consumed, and a main peak with the desired mass number was detected. The reaction mixture was quenched with H2O (30.0 mL) at 0°C and then extracted with DCM (20.0 mL×3). The combined organic layers were washed with brine (30.0 mL), dried over Na2SO4, filtered and concentrated under reduced pressure to give compound 3 (607 mg, 1.73 mmol, crude product) as a yellow oil.

[0159] LCMS: m / z=352.0 (M+H) + , Rt=1.785min.

[0160] Compound 3 (607 mg, 1.73 mmol) was dissolved in toluene (6.00 mL). 3a (443 mg, 1.73 mmol) was then added. The mixture was stirred at 110°C for 2 hours. LCMS (Rt = 2.157 min) showed that compound 3 was completely consumed, with a major peak with the desired mass detected. The reaction mixture was filtered and concentrated under reduced pressure to afford compound 4 (887 mg, 1.73 mmol, crude) as a yellow oil.

[0161] LCMS: m / z=514.0 (M+H) + , Rt=2.157min.

[0162] POCl3 (529 mg, 3.45 mmol, 321 μL) was dissolved in toluene (8.00 mL), followed by the addition of compound 4 (887 mg, 1.73 mmol) and DIEA (446 mg, 3.45 mmol, 601 μL). The mixture was stirred at 110°C for 2 hours. LCMS (Rt = 2.279 min) showed that compound 4 was completely consumed, with a major peak with the desired mass detected. The reaction mixture was quenched with H2O (20.0 mL) at 20°C, followed by extraction with ethyl acetate (10.0 mL x 3). The combined organic layers were washed with brine (10.0 mL), dried over Na2SO4, filtered, and concentrated under reduced pressure to afford compound 5 (918 mg, 1.72 mmol, crude) as a yellow solid.

[0163] LCMS: m / z=532.0 (M+H) + , Rt=2.279min.

[0164] Compound 5 (918 mg, 1.72 mmol) was dissolved in DCM (10.0 mL). Compound 5a (415 mg, 2.59 mmol) and TEA (872 mg, 8.62 mmol, 1.20 mL) were then added. The mixture was stirred at 25°C for 2 hours. LCMS (Rt = 4.686 min) showed that compound 5 was completely consumed, and a major peak with the desired mass was detected. The reaction mixture was filtered and concentrated under reduced pressure to obtain a residue. The residue was purified by preparative HPLC (TFA conditions) (column: Phenomenex luna C18 150 × 40 mm × 15 μm; mobile phase: [H2O(TFA)-ACN]; gradient: 44%-74% B in 10.0 min). The product was then purified by SFC (chromatographic column: DAICEL CHIRALPAK AD (250 mm × 30 mm, 10 μm); mobile phase: [CO2-EtOH (0.1% NH3H2O)]; B%: 40%, isocratic elution mode) to obtain compound Q4-P1 (40.0 mg, purity 98.35%) as a white solid, and compound Q4-P2 (38.80 mg, purity 95.56%) as a white solid.

[0165] LCMS: m / z=620.1 (M+H) + , Rt=4.686min.

[0166] Compound Q4-P1:

[0167] LCMS: m / z=620.0 (M+H) + , Rt=3.416min.

[0168] 1 H NMR: (400MHz, CDCl3-d)δ7.55(br d,J=8.6Hz,3H),7.48-7.53(m,1H),7.26-7.37(m,6H),7.14(d,J=6.9Hz,2H),4.85-5.00(m, 2H), 4.75 (dd, J=11.4, 5.4Hz, 1H), 4.56-4.65 (m, 1H), 4.13-4.24 (m, 3H), 3.49-3.57 (m, 2H).

[0169] Compound Q4-P2:

[0170] LCMS: m / z=620.0 (M+H) + , Rt=3.415min.

[0171] 1H NMR: (400MHz, CDCl3-d) δ7.52-7.63(m,3H),7.50(br d,J=3.8Hz,1H),7.26-7.37(m,6H),7.14(br d,J=7.4Hz,2H),4.88-5.11(m,2H),4.74(dd,J=11.4,5.3Hz,1H),4.60(t,J=11.3Hz,1H),4.12-4.23(m,3H),3.50-3.57(m,2H).

[0172] Preparation Example 21: Synthesis of (Z)-3-(4-chlorophenyl)-4-phenyl-N-(2-sulfamoylethyl)-N'-((5-(trifluoromethyl)furan-2-yl)sulfonyl)-4,5-dihydro-1H-pyrazole-1-carboximide (S, R enantiomers of Compound Q5)

[0173] To a solution of compound 1 (1.00 g, 6.00 mmol, 1.00 eq.) in DCM (10.0 mL) were added TEA (1.82 g, 18.0 mmol, 2.51 mL, 3.00 eq.) and compound 1a (571 mg, 7.80 mmol, 820 μL, 1.30 eq.). The reaction mixture was stirred at 25° C. for 4 hours. TLC (petroleum ether:ethyl acetate=3:1, R f =0.59) indicated that compound 1 was completely consumed and a new spot was formed. The reaction was judged to be complete according to the TLC results. The reaction mixture was filtered and concentrated under reduced pressure to obtain a residue. The residue was purified by flash silica gel chromatography ( 12g The product was purified by silica gel flash column (0-25% ethyl acetate / petroleum ether gradient @ 30 mL / min) to afford compound 2 as a colorless oil (1.00 g, yield 77.9%, purity 95%).

[0174] LCMS: m / z=221.1 (M+NH3+H) + , Rt=1.858min.

[0175] 1 H NMR: (400MHz, CDCl3-d) δ7.55 (s, 1H), 7.00 (d, J = 3.4Hz, 1H), 6.49 (br d, J = 1.5Hz, 1H), 4.46-4.70 (m, 1H), 1.27 (s, 9H).

[0176] To a 15.0 mL reaction vial equipped with a stir bar was added a solution of compound 2 (850 mg, 4.18 mmol, 1.00 eq.), Ru(bpy)3Cl2·6H2O (25.0 mg, 33.4 μmol, 0.00800 eq.), and 1-oxo-4-phenylpyridin-1-ium (1.43 g, 8.36 mmol, 2.00 eq.). The vial was sealed and placed under nitrogen. (2,2,2-trifluoroacetyl) 2,2,2-trifluoroacetate (2.20 g, 10.4 mmol, 1.45 mL, 2.50 eq.) was then added. The reaction was stirred and illuminated with a 10 W blue LED lamp (3 cm away). The reaction temperature was maintained at 25°C with cooling water for 14 hours. TLC (petroleum ether:ethyl acetate = 10:1, R f =0.29) indicated that compound 2 had reacted completely and formed multiple new spots. The reaction was judged to be complete according to TLC results. The reaction mixture was filtered and concentrated under reduced pressure to obtain a residue. The residue was purified by flash silica gel chromatography ( 4g The product was purified by silica gel flash column with 0-25% ethyl acetate / petroleum ether gradient at 18 mL / min as eluent to afford compound 3 as a brown oil (672 mg, 53.3% yield, 90% purity).

[0177] LCMS: m / z=289.0 (M+NH3+H) + , Rt=2.382min.

[0178] 1 H NMR: (400MHz, CDCl3-d) δ7.01-7.06(m,1H),6.86-6.92(m,1H),4.62-4.79(m,1H),1.31(s,9H).

[0179] 19 F NMR: (376MHz, CDCl3-d)δ-64.339(s,3F).

[0180] A mixture of compound 3 (250 mg, 922 μmol, 1.00 eq.) and TFA (1.63 g, 14.3 mmol, 1.06 mL, 15.5 eq.) dissolved in DCM (0.250 mL) was degassed and purged with N 2 three times, and then the mixture was stirred at 25 ° C. under N 2 atmosphere for 14 hours. LCMS (Rt = 1.504 min) showed that compound 3 was completely consumed, and a major peak with the desired mass was detected. The reaction mixture was filtered and concentrated under reduced pressure to give 4 (198 mg, crude product) as a yellow oil.

[0181] LCMS: m / z=215.9 (M+H) + , Rt=1.504min.

[0182] Compound 4 (198 mg, 920 μmol, 1.00 eq.) and TEA (233 mg, 2.30 mmol, 320 μL, 2.50 eq.) were dissolved in DCM (2.00 mL) at 0°C. Compound 4a (173 mg, 1.10 mmol, 138 μL, 1.20 eq.) was then added. The mixture was stirred at 0°C for 4 hours. LCMS (Rt = 2.165 min) showed that compound 4 was completely consumed, and a major peak with the desired mass was detected. The reaction mixture was concentrated under reduced pressure to give compound 5 (308 mg, crude product) as a brown oil.

[0183] LCMS: m / z=336.0 (M+H) + , Rt=2.165min.

[0184] Compound 5 (308 mg, 919 μmol, 1.00 eq.) was dissolved in toluene (3.00 mL). Compound 5a (236 mg, 919 μmol, 1.00 eq.) was then added. The mixture was stirred at 110° C. for 1 hour. LCMS (Rt=2.885 min) showed that compound 5 was completely consumed, with a major peak with the desired mass detected. The reaction mixture was concentrated under reduced pressure to give compound 6 (457 mg, crude) as a brown oil.

[0185] LCMS: m / z=497.9 (M+H) + , Rt=2.885min.

[0186] Compound 6 (457 mg, 918 μmol, 1.00 eq.) was dissolved in toluene (3.00 mL). POCl (281 mg, 1.84 mmol, 171 μL, 2.00 eq.) and DIEA (237 mg, 1.84 mmol, 320 μL, 2.00 eq.) were then added. The mixture was stirred at 100° C. for 2 hours. LCMS (Rt=3.039 min) showed that compound 6 was completely consumed, and a main peak with the desired mass number was detected. The reaction mixture was quenched with 1 M NaOH (20.0 mL) at 20° C. and then extracted with ethyl acetate (10.0 mL×3). The combined organic layers were washed with brine (10.0 mL), dried over NaSO, filtered, and concentrated under reduced pressure to give compound 7 (474 ​​mg, crude product) as a brown oil.

[0187] LCMS: m / z=516.1 (M+H) + , Rt=3.309min.

[0188] Compound 7 (473 mg, 916 μmol, 1.00 eq.) was dissolved in DCM (3.00 mL). TEA (464 mg, 4.58 mmol, 638 μL, 5.00 eq.) and compound 7a (221 mg, 1.37 mmol, 1.50 eq.) were then added. The mixture was stirred at 25°C for 4 hours. LCMS (Rt = 2.646 min) showed complete consumption of compound 7, with a major peak of the desired mass detected. The reaction mixture was filtered and concentrated under reduced pressure to obtain a residue. The residue was purified by preparative HPLC (TFA conditions) (column: CD24-XPT C18 150 × 25 mm × 7 μm; mobile phase: [H2O(TFA)-ACN]; gradient: 42% to 62% B over 10.0 min) to obtain compound 8 (300 mg, 54.2% yield, 99.9% purity) as a white solid.

[0189] LCMS: m / z=604.1 (M+H) + , Rt=2.646min.

[0190] LCMS: m / z=604.0 (M+H) + , Rt=2.005min.

[0191] Compound 8 was purified by SFC: chromatographic column: DAICEL CHIRALPAK AD (250 mm × 30 mm, 10 μm); mobile phase: [CO2-EtOH (0.1% NH3H2O)]; B%: 45%, isocratic elution mode to obtain compound Q5-P1 (60.0 mg, purity 99.9%, 99.9% ee) as a white solid, and compound Q5-P2 (90.0 mg, yield 99.9%, 97.8% ee) as a white solid.

[0192] LCMS: m / z=614.1 (M+H) + , Rt=2.647min.

[0193] Compound Q5-P1:

[0194] LCMS: m / z=614.3 (M+H) + , Rt=1.997min.

[0195] HPLC: Rt = 4.267 min.

[0196] SFC: Rt = 1.628 min.

[0197] 1 H NMR: (400MHz, CDCl3-d) δ7.66-7.80(m,1H),7.58(d,J=8.6Hz,2H),7.27-7.39(m,5H),7.14(d,J=6.9Hz,2H),6.94(d,J= 3.4Hz,1H),6.76-6.85(m,1H),5.13(s,2H),4.73-4.84(m,1H),4.58-4.70(m,1H),4.00-4.29(m,3H),3.45-3.66(m,2H).

[0198] 19 F NMR: (376MHz, CDCl3-d)δ-64.159 (s, 3F).

[0199] Compound Q5-P2:

[0200] LCMS: m / z=614.3 (M+H) + , Rt=2.001min.

[0201] HPLC: Rt = 4.273 min.

[0202] SFC: Rt = 2.060 min.

[0203] 1 H NMR: (400MHz, CDCl3-d)δ7.65(br t,J=5.6Hz,1H),7.57(d,J=8.5Hz,2H),7.27-7.38(m,5H),7.07-7.21(m,2H),6.93(d,J=3.4Hz,1H),6.76-6.84(m,1H),4.94(br s,2H),4.77(dd,J=11.4,5.3Hz,1H),4.58-4.71(m,1H),4.08-4.30(m,3H),3.48-3.60(m,2H).

[0204] 19 F NMR: (376MHz, CDCl3-d)δ-64.166 (s, 3F).

[0205] Preparation Example 22: Synthesis of S and R enantiomers of compound Q2

[0206] Compound 1 (5.00 g, 22.2 mmol) and TEA (5.62 g, 55.5 mmol, 7.73 mL) were dissolved in DCM (50.0 mL) at 0°C. Compound 1a (3.48 g, 22.2 mmol, 2.79 mL) was then added dropwise at 0°C. The mixture was stirred at 0°C for 4 hours. LCMS (Rt = 1.804 min) showed that compound 1 was completely consumed, and a main peak with the desired mass number was detected. The reaction mixture was quenched with H2O (30.0 mL) at 0°C and then extracted with DCM (20.0 mL × 3). The combined organic layers were washed with brine (30.0 mL), dried over Na2SO4, filtered and concentrated under reduced pressure to obtain compound 2 (7.65 g, 22.1 mmol, crude product) as a white solid.

[0207] LCMS: m / z=346.1 (M+H) + , Rt=1.804min.

[0208] Compound 2 (3.00 g, 8.69 mmol) was dissolved in toluene (3.00 mL). Compound 2a (2.23 g, 8.69 mmol) was then added. The mixture was stirred at 110° C. for 2 hours. LCMS (Rt=1.688 min) showed that compound 2 was completely consumed, with a major peak with the desired mass detected. The reaction mixture was filtered and concentrated under reduced pressure to afford compound 3 (4.41 g, 8.68 mmol, crude) as a yellow oil.

[0209] LCMS: m / z=508.2 (M+H) + , Rt=1.804min.

[0210] POCl3 (603 mg, 3.94 mmol, 367 μL) was dissolved in toluene (10.0 mL). Compound 3 (1.00 g, 1.97 mmol) and DIEA (508 mg, 3.94 mmol, 685 μL) were then added. The mixture was stirred at 110°C for 2 hours. LCMS (Rt = 1.783 min) showed that compound 3 was completely consumed, and a main peak with the required mass number was detected. The reaction mixture was quenched with H2O (40.0 mL) at 20°C, and then extracted with ethyl acetate (20.0 mL×3). The combined organic layers were washed with brine (20.0 mL), dried over Na2SO4, filtered and concentrated under reduced pressure to obtain compound 3 (1.01 g, crude product) as a yellow oil.

[0211] LCMS: m / z=526.2 (M+H) +, Rt=1.783min.

[0212] To a solution of compound 5 (10.0 g, 88.0 mmol, HCl salt) in toluene (50.0 mL) were added pyridine (13.9 g, 176 mmol, 14.2 mL) and compound 5a (19.5 g, 132 mmol). The mixture was stirred at 110° C. for 3 hours. LCMS (Rt=1.942 min) showed that compound 5 was completely consumed, and a major peak with the desired mass was detected. The reaction mixture was filtered and concentrated under reduced pressure to obtain a residue. The residue was purified by flash silica gel chromatography ( 80g Purification was performed by silica gel flash chromatography (eluent: 0-30% ethyl acetate / petroleum ether gradient @ 50 mL / min) to obtain compound 6 as a white solid (15.0 g, 65.1 mmol, yield 74.0%, purity 90.0%).

[0213] LCMS: m / z=208.1 (M+H) + , Rt=1.942min.

[0214] 1 H NMR: (400MHz, CDCl3-d) δ7.82-7.90 (m, 2H), 7.69-7.77 (m, 2H), 3.88 (t, J = 7.1Hz, 2H), 2.74-2.98 (m, 2H), 1.43 (t, J = 8.6Hz, 1H).

[0215] To a solution of compound 6 (1.29 g, 6.22 mmol) in DCM (10.0 mL) was added Ac2O (635 mg, 6.22 mmol, 584 μL) and sulfonyl chloride (1.68 g, 12.4 mmol, 1.24 mL) at -20°C. The mixture was stirred at 25°C for 1 hour. TLC (petroleum ether:ethyl acetate = 1:1, R f=0.25) showed that compound 6 was completely consumed and a new spot was formed. The reaction mixture was filtered and concentrated under reduced pressure to give the product. TEA (3.14 g, 31.0 mmol, 4.32 mL) was added to a solution of bis(2,4-dimethoxybenzyl)amine (1.97 g, 6.21 mmol) in DCM (16.0 mL) at 0 ° C and N2. A solution of the crude product in DCM (8.00 mL) was then added dropwise at 0 ° C and N2. The mixture was stirred at 0 ° C for 1 hour. LCMS (Rt = 2.573 min) showed that the residue was completely consumed and a main peak with the desired mass number was detected. The reaction mixture was filtered and concentrated under reduced pressure to give a residue. The residue was purified by flash silica gel chromatography ( 40g The product was purified by silica gel flash chromatography (0-60% ethyl acetate / petroleum ether gradient @ 60 mL / min) to afford compound 7 (1.51 g, 2.16 mmol, 34.7% yield, 77.0% purity) as a yellow oil.

[0216] LCMS: m / z=539.2 (M+H) + , Rt=2.573min.

[0217] 1 H NMR: (400MHz, CDCl3-d) δ7.80-7.88(m,2H),7.68-7.75(m,2H),7.15(d,J=9.0Hz,2H),6.37-6.43(m,4H),4.36(d,J=15.3Hz ,2H),4.11(d,J=15.3Hz,2H),3.90-3.98(m,1H),3.83-3.89(m,1H),3.78(s,6H),3.76(s,6H),3.11(tt,J=13.4,6.6Hz,2H).

[0218] To a solution of compound 7 (500 mg, 928 μmol) in MeOH (2.00 mL) were added (NH4)2CO3 (356 mg, 3.71 mmol, 396 μL) and PIFA (1.20 g, 2.78 mmol). The mixture was stirred at 25°C for 1 hour. LCMS (Rt = 2.531 min) showed that compound 7 was completely consumed, and a major peak with the desired mass was detected. The reaction mixture was filtered and concentrated under reduced pressure to obtain a residue. The residue was purified by flash silica gel chromatography ( 4g The product was purified by silica gel flash chromatography (0-55% ethyl acetate / petroleum ether gradient @ 25 mL / min) to afford compound 8 (105 mg, 189 μmol, crude product) as a white solid.

[0219] LCMS: m / z=554.2 (M+H) + , Rt=2.531min.

[0220] Compound 8 (105 mg, 189 μmol) and N₂H₄·H₂O (80.0 mg, 1.60 mmol, 77.5 μL, 100% purity) were dissolved in MeOH (2.00 mL). The reaction mixture was stirred at 25°C under N₂ for 1 hour. LCMS (Rt = 1.885 min) showed complete consumption of compound 8, with a major peak of the desired mass detected. The reaction mixture was concentrated under reduced pressure to afford compound 9 (80.0 mg, crude) as a white solid.

[0221] LCMS: m / z=424.2 (M+H) + , Rt=1.885min.

[0222] Compound 4 (99.4 mg, 188 μmol) was dissolved in DCM (2.00 mL). TEA (95.5 mg, 944 μmol, 131 μL) and compound 9 (80.0 mg, 188 μmol) were then added. The mixture was stirred at 25°C for 2 hours. LCMS (Rt = 3.219 min) showed that compound 9 was completely consumed and a major peak with the desired mass was detected. The reaction mixture was filtered and concentrated under reduced pressure to obtain a residue. The residue was purified by high-speed silica gel chromatography ( 4g The product was purified by flash chromatography on silica gel with a gradient of 0-45% ethyl acetate / petroleum ether at 25 mL / min as eluent to afford compound 10 (80.0 mg, 87.58 μmol, 46.3% yield) as a brown oil.

[0223] LCMS: m / z = 913.2 (M+H) + , Rt=3.219min.

[0224] Compound 10 (50 mg, 54.7 μmol) was dissolved in TFA (2.00 mL) and MsOH (0.40 mL). The mixture was stirred at 25°C for 12 hours. LCMS (Rt = 2.488 min) showed that compound 10 was completely consumed, and a major peak with the desired mass was detected. The reaction mixture was filtered and concentrated under reduced pressure to obtain a residue. The residue was purified by rapid silica gel chromatography ( 4g The product was purified by silica gel flash chromatography (0-75% ethyl acetate / petroleum ether gradient @ 20 mL / min as eluent) to afford compound 11 as a white solid (13.0 mg, 34.8% yield, 90% purity).

[0225] LCMS: m / z=613.2 (M+H) + , Rt=2.488min.

[0226] LCMS: m / z=613.2 (M+H) + , Rt=2.486min.

[0227] Compound 11 (13.0 mg) was purified by SFC: chromatographic column: DAICEL CHIRALPAK AD (250 mm × 30 mm, 10 μm); mobile phase: [CO2-EtOH (0.1% NH3H2O)]; B%: 55%, isocratic elution mode to obtain compound Q2-P1 (4.20 mg, purity 98.70%) as a beige solid, and compound Q2-P2 (4.10 mg, yield 94.55%) as a beige solid.

[0228] Compound Q2-P1:

[0229] LCMS: m / z=613.2 (M+H) + , Rt=2.504min.

[0230] Compound Q2-P2:

[0231] LCMS: m / z=613.3 (M+H) + , Rt=2.507min.

[0232] Preparation Example 23: Synthesis of (Z)-3-(5-cyanothiophene-2-yl)-4-phenyl-N-(2-sulfamoylethyl)-N'-((4-(trifluoromethyl)phenyl)sulfonyl)-4,5-dihydro-1H-pyrazole-1-carboximide (S, R enantiomers of compound Q6)

[0233] To a solution of compound 1 (11.9 g, 72.8 mmol, 7.05 mL, 1.50 eq.) and compound 1a (7.50 g, 48.5 mmol, 6.47 mL, 1.00 eq.) in DCM (50.0 mL) was added AlCl₃ (9.70 g, 72.8 mmol, 3.98 mL, 1.50 eq.) at 0°C. The reaction mixture was stirred at 25°C under a N₂ atmosphere for 1 hour. TLC (petroleum ether:ethyl acetate = 10:1, R f =0.60) indicated that compound 1 was completely consumed and two new spots were formed. The reaction mixture was quenched with 1.00M HCl (30.0 mL) and extracted with DCM (30.0 mL×3). The combined organic layers were washed with brine (30.0 mL×2), dried over Na2SO4, filtered and concentrated under reduced pressure to give a residue. The residue was purified by flash silica gel chromatography ( 20g The product was purified by silica gel flash column with 0-4% ethyl acetate / petroleum ether gradient at 25 mL / min as eluent to afford compound 2 (7.24 g, 53.1% yield) as a brown solid.

[0234] 1 H NMR: (400MHz, CDCl3-d) δ7.42-7.56(m,1H),7.27-7.38(m,5H),7.03-7.15(m,1H),4.13(s,2H).

[0235] To a solution of compound 2 (4.50 g, 16.0 mmol, 1.00 eq.) in H 2 O (40.0 mL) and THF (8.00 mL) were added Zn (CN) 2 (2.16 g, 18.4 mmol, 1.17 mL, 1.15 eq.) and tBuXPhos Pd G 3 (1.27 g, 1.60 mmol, 0.100 eq.). The reaction mixture was stirred at 25 ° C. under N 2 atmosphere for 14 hours. TLC (petroleum ether: ethyl acetate = 10: 1, R f =0.06) indicated that compound 2 was completely consumed and a new spot was formed. The reaction was judged to be complete according to the TLC result. The reaction mixture was quenched with 30.0 mL of NaClO aqueous solution at 25 ° C, and then extracted with 90.0 mL (30.0 mL×3) of ethyl acetate. The combined organic layer was washed with 90.0 mL (30.0 mL×3) of brine, dried over Na2SO4, filtered and concentrated under reduced pressure to obtain a residue. The residue was purified by flash silica gel chromatography ( 40g The product was purified by silica gel flash column (0-9% ethyl acetate / petroleum ether gradient @ 30 mL / min as eluent) to afford brown solid compound 3 (3.12 g, yield 77.2%, purity 90.0%).

[0236] LCMS: m / z=228.1 (M+H) + , Rt=2.400min.

[0237] 1 H NMR: (400MHz, CDCl3-d) δ7.69 (d, J=4.0Hz, 1H), 7.60 (d, J=4.0Hz, 1H), 7.24-7.43 (m, 5H), 4.22 (s, 2H).

[0238] To a solution of compound 3 (3.12 g, 13.7 mmol, 1.00 eq.) and formaldehyde (5.57 g, 68.6 mmol, 5.11 mL, 5 eq.) in MeOH (25.0 mL) were added piperidine (1.17 g, 13.7 mmol, 1.36 mL, 1.00 eq.) and acetic acid (1.65 g, 27.4 mmol, 1.57 mL, 2.00 eq.). The mixture was stirred at 70° C. for 2 hours. TLC (petroleum ether: ethyl acetate = 10:1, R f =0.32) indicated that compound 3 was completely consumed and a new spot was formed. The reaction was judged to be complete according to TLC results. The reaction mixture was concentrated under reduced pressure to obtain compound 4 (3.28 g, crude product) as a yellow oil.

[0239] LCMS: m / z=240.1 (M+H) + , Rt=2.549min.

[0240] Compound 4 (1.30 g, 5.43 mmol, 1.00 eq.) and hydrazine hydrate (810 mg, 16.2 mmol, 785 μL, 2.98 eq.) were dissolved in EtOH (10.0 mL). The mixture was stirred at 80° C. for 1 hour. LCMS (Rt=1.092 min) showed that compound 4 was completely consumed, and a major peak with the desired mass was detected. The reaction mixture was concentrated under reduced pressure to obtain compound 5 (1.40 g, crude product) as a yellow solid.

[0241] LCMS: m / z=254.0 (M+H) + , Rt=1.092min.

[0242] Compound 5a (1.10 g, 3.19 mmol, 1.00 eq.) was dissolved in toluene (2.00 mL). Compound 5 (807.01 mg, 3.19 mmol, 1.00 eq.) was then added. The mixture was stirred at 110°C for 2 hours. LCMS (Rt = 2.765 min) showed complete consumption of compound 5, with a major peak of the desired mass detected. The reaction mixture was filtered and concentrated under reduced pressure to obtain a residue. The residue was purified by preparative HPLC (column: CD18-Welch Ultimate C18 150 × 40 × 7 μm; mobile phase: [H2O (0.05% HCl)-ACN]; gradient: 51% to 81% B over 15.0 min) to obtain compound 6 (430 mg, 21.4% yield, 80.0% purity) as a yellow solid.

[0243] LCMS: m / z=504.9 (M+H) + , Rt=2.765min.

[0244] LCMS: m / z=504.5 (M+H) + , Rt=2.873min.

[0245] 1 H NMR: (400MHz, CDCl3-d)δ8.68(s,1H),8.32(d,J=8.6Hz,2H),7.85(d,J=8.0Hz,2H),7.33-7.45(m,5H ),7.16-7.18(m,1H),6.93(d,J=4.0Hz,1H),4.62-4.74(m,1H),4.35-4.49(m,1H),3.87-4.00(m,1H).

[0246] 19 F NMR: (376MHz, CDCl3-d)δ-63.239(s,3F).

[0247] Compound 6 (120 mg, 238 μmol, 1.00 eq.) was dissolved in toluene (5.00 mL). POCl (72.9 mg, 476 μmol, 44.3 μL, 2.00 eq.) and DIEA (61.5 mg, 476 μmol, 82.9 μL, 2.00 eq.) were then added. The mixture was stirred at 110° C. for 2 hours. LCMS (Rt=2.929 min) showed that compound 6 was completely consumed, and a major peak with the desired mass was detected. The reaction mixture was quenched with 1 M NaOH (20.0 mL) at 20° C. and then extracted with ethyl acetate (10.0 mL×3). The combined organic layers were washed with brine (10.0 mL), dried over NaSO, filtered, and concentrated under reduced pressure to obtain compound 7 (124 mg, crude product) as a brown oil.

[0248] LCMS: m / z=522.9 (M+H) + , Rt=2.929min.

[0249] Compound 7 (110 mg, 210 μmol, 1.00 eq.) was dissolved in DCM (2 mL). TEA (106 mg, 1.05 mmol, 146 μL, 5.00 eq.) and 2-aminoethanesulfonamide hydrochloride (50.7 mg, 316 μmol, 1.50 eq.) were then added. The mixture was stirred at 25 ° C for 2 hours. TLC (petroleum ether: ethyl acetate = 1:2, product: R f =0.46) indicated that compound 7 was completely consumed and many new spots were formed. The reaction was determined to be complete based on TLC results. The reaction mixture was filtered and concentrated under reduced pressure to obtain a residue. The residue was purified by preparative TLC (SiO2, petroleum ether / ethyl acetate = 1 / 2) to obtain compound 8 (50.0 mg, 38.5% yield, 99.0% purity) as a white solid.

[0250] LCMS: m / z=611.2 (M+H) + , Rt=1.910min.

[0251] HPLC: Rt = 2.062 min.

[0252] 1H NMR: (400MHz, CDCl3-d)δ8.05(d,J=8.3Hz,2H),7.67-7.75(m,2H),7.54-7.64(m,1 H),7.30-7.44(m,4H),7.19(dd,J=7.7,1.6Hz,2H),6.92(d,J=4.0Hz,1H),5.19(br s,2H),4.57-4.74(m,2H),4.07-4.28(m,3H),3.48-3.60(m,2H).

[0253] 19 F NMR: (376MHz, CDCl3-d)δ-62.951 (s, 3F).

[0254] Compound 8 was purified by SFC: chromatographic column: DAICEL CHIRALPAK AD (250 mm × 30 mm, 10 μm); mobile phase: [CO2-EtOH (0.1% NH3H2O)]; B%: 50%, isocratic elution mode to obtain compound Q6-P1 (15.0 mg, yield 35.8%, purity 95.4%) as a white solid, and compound Q6-P2 (16.0 mg, yield 39.28%, purity 98.2%) as a white solid.

[0255] LCMS: m / z=610.5 (M+H) + , Rt=2.639min.

[0256] Compound Q6-P1:

[0257] LCMS: m / z=611.1 (M+H) + , Rt=1.447min.

[0258] HPLC: Rt=3.740 min.

[0259] SFC: Rt = 1.901 min.

[0260] 1H NMR: (DMSO-d6, 400MHz) δ8.12-8.25 (m, 1H), 8.04 (d, J = 8.4Hz, 2H), 7.77-7.91 (m, 3H), 7.34-7.41 (m, 2H), 7. 27-7.33(m,3H),7.22-7.26(m,1H),7.03(s,2H),4.99-5.12(m,1H),4.66(s,1H),4.05-4.19(m,1H),3.74(br d,J=5.3Hz,2H),3.26-3.30(m,2H).

[0261] 19 F NMR: (DMSO-d6,377MHz)δ-61.363(s,3F).

[0262] Compound Q6-P2:

[0263] LCMS: m / z=611.0 (M+H) + , Rt=1.773min.

[0264] HPLC: Rt=3.746 min.

[0265] SFC: Rt = 2.609 min.

[0266] 1 H NMR: (400MHz, CDCl3-d)δ8.05(d,J=8.3Hz,2H),7.72(d,J=8.4Hz,2H),7.52(br d,J=5.9Hz,1H),7.33-7.44(m,4H),7.20(br d,J=6.4Hz,2H),6.93(d,J=4.0Hz,1H),4.94(s,2H),4.55-4.75(m,2H),4.07-4.26(m,3H),3.52(t,J=5.9Hz,2H).

[0267] 19 F NMR: (376MHz, CDCl3-d) δ-62.954 (s, 3F).

[0268] Preparation Example 24: Synthesis of (Z)-3-(5-cyanothiophene-2-yl)-4-phenyl-N-((R)-2-sulfamoylpropyl)-N'-((4-(trifluoromethyl)phenyl)sulfonyl)-4,5-dihydro-1H-pyrazole-1-carboximide (S, R enantiomers of compound Q7)

[0269] To a solution of compound 1 (25.0 g, 333 mmol, 26.2 mL, 1.00 eq.) in toluene (250 mL) was added Et3N (3.37 g, 33.3 mmol, 4.63 mL, 0.100 eq.) and compound 1a (49.3 g, 333 mmol, 1.00 eq.). The mixture was stirred at 130 ° C for 3 hours. LC-MS (Rt = 2.334 min) showed that compound 1 was completely consumed and a peak of the desired mass was detected. The reaction mixture was filtered and concentrated under reduced pressure to give compound 2 (68.3 g, crude product) as a white solid.

[0270] LCMS: m / z=206.1 (M+H) + , Rt=2.334min.

[0271] To a solution of compound 2 (68.3 g, 333 mmol, 1.00 eq.) in DCM (250 mL) at 0°C were added TEA (101 g, 998 mmol, 139 mL, 3.00 eq.) and DMAP (407 mg, 3.33 mmol, 0.0100 eq.), followed by 4-methylbenzenesulfonyl chloride (69.8 g, 366 mmol, 1.10 eq.). The mixture was stirred at 25°C for 10 hours. LC-MS (Rt = 2.331 min) showed that compound 2 was completely consumed, and a peak of the desired mass was detected. The reaction mixture was filtered and concentrated under reduced pressure to obtain a residue. The crude product was triturated with 50.0 mL of (PE:EA = 10:1) at 25°C for 30 min to obtain compound 3 (89.0 g, 74.4% yield) as a white solid.

[0272] LCMS: m / z=360.1 (M+H) + , Rt=2.331min.

[0273] Compound 3 (38.1 g, 106 mmol, 1.00 eq.) and compound 3a (13.1 g, 117 mmol, 1.10 eq.) were added to DMF (200 mL), followed by anhydrous KCO (29.3 g, 212 mmol, 2.00 eq.), and the mixture was stirred at 60°C for 12 hours. LC-MS (Rt = 1.618 min) showed the detection of the desired compound. The reaction mixture was extracted with 180 mL of EA (60.0 mL x 3). The combined organic layers were washed with 180 mL of water (60.0 mL x 3), dried over NaSO, filtered, and concentrated under reduced pressure to obtain a residue. The residue was purified by preparative HPLC (chromatographic column: Ultimate XB-SIO2 250×50 mm×10 μm; mobile phase: [hexane-EtOH]; gradient: 1%-26% B in 15.0 min) to give white solid compound 4 (20.0 g, yield 63.0%).

[0274] LCMS: m / z=299.8 (M+H) + , Rt=1.618min.

[0275] Compound 4 (8.30 g, 27.7 mmol, 1.00 eq.) was dissolved in DCM (15.0 mL). mCPBA (8.44 g, 41.6 mmol, 1.50 eq.) was then added at 0°C. The mixture was stirred at 25°C for 10 hours. TLC (petroleum ether:ethyl acetate = 1:1, R f =0.08) indicated that compound 4 was completely consumed and multiple new spots appeared. The reaction mixture was quenched with saturated aqueous Na2S2O3 solution (10 mL) and extracted with EtOAc (10.0 mL×2). The combined organic layers were washed with saturated aqueous NaHCO3 solution (10.0 mL) and brine (10.0 mL), dried over Na2SO4, filtered and concentrated under reduced pressure. The residue was purified by flash silica gel chromatography ( 40g The product was purified by silica gel flash column (eluting with 0-55% ethyl acetate / petroleum ether gradient @ 40 mL / min) to afford compound 5 (8.89 g, 96.7% yield) as a white solid.

[0276] 1H NMR: (400MHz, CDCl3-d) δ8.95 (d, J = 4.9Hz, 2H), 7.79-7.88 (m, 2H), 7.70-7.77 (m, 2H) ,7.55(t,J=4.8Hz,1H),4.23-4.40(m,2H),3.97-4.10(m,1H),1.51(d,J=7.0Hz,3H).

[0277] A solution of compound 5 (8.00 g, 24.1 mmol, 1.00 eq.) dissolved in MeOH (40.0 mL) was cooled to 0° C. and treated with sodium methoxide (5.00 M, 5.79 mL, 1.20 eq.). The mixture was stirred at this temperature for 2 hours, then treated with a solution of solid sodium acetate (2.97 g, 36.2 mmol, 1.50 eq.) and hydroxylamine sulfonic acid (4.10 g, 36.2 mmol, 1.50 eq.) dissolved in H O (40 mL), heated to 20° C. and maintained for 12 hours. TLC (petroleum ether: ethyl acetate = 1:1, product: R f =0.33) indicated that compound 5 was completely consumed and a new spot was formed. The reaction mixture was filtered and concentrated under reduced pressure to obtain a residue. The residue was purified by flash silica gel chromatography ( 40g The product was purified by silica gel flash column (eluting with 0-50% ethyl acetate / petroleum ether gradient @ 45 mL / min) to afford compound 6 (2.63 g, 40.5% yield) as a white solid.

[0278] 1 H NMR: (400MHz, DMSO-d6) δ7.77-8.06 (m, 4H), 7.01 (s, 2H), 4.02 (dd, J = 14.0, 5.1 Hz, 1H), 3.72 (dd, J = 13.9, 9.7Hz, 1H), 3.37-3.42 (m, 1H), 1.21 (d, J = 6.9Hz, 3H).

[0279] Compound 6 (1.40 g, 5.22 mmol, 1.00 eq.) and NH2NH2·H2O (900 mg, 18.0 mmol, 872 μL, 100% purity, 3.45 eq.) were dissolved in EtOH (15.0 mL). The mixture was stirred at 80°C under N2 for 1 hour. LCMS (Rt = 0.410 min) showed complete consumption of compound 6, with a major peak of the desired mass detected. The reaction mixture was concentrated under reduced pressure to obtain a residue. The residue was purified by preparative HPLC (column: CD01-Phenomenex Luna C18 150 × 25 mm × 10 μm; mobile phase: [H2O (0.225% FA)-ACN]; gradient: 0% to 30% B over 10.0 min) to obtain compound 7 (1.20 g, 99.8% yield) as a colorless oil.

[0280] LCMS: m / z=139.1 (M+H) + , Rt=0.410min.

[0281] 1 H NMR: (400MHz, DMSO-d6) δ3.03-3.10 (m, 2H), 2.69-2.81 (m, 1H), 1.26 (d, J = 6.8Hz, 3H).

[0282] To a solution of compound 1a (11.9 g, 72.8 mmol, 7.05 mL, 1.50 eq.) and compound 1aa (7.50 g, 48.5 mmol, 6.47 mL, 1.00 eq.) in DCM (50.0 mL) was added AlCl₃ (9.70 g, 72.8 mmol, 3.98 mL, 1.50 eq.) at 0°C. The reaction mixture was stirred at 25°C under a N₂ atmosphere for 1 hour. TLC (petroleum ether:ethyl acetate = 10:1, R f =0.60) showed that compound 1a was completely consumed and two new spots were formed. The reaction mixture was quenched with 1.00M HCl (30.0mL) and extracted with DCM (30.0mL×3). The combined organic layers were washed with brine (30.0mL×2), dried over Na2SO4, filtered and concentrated under reduced pressure to give a residue. The residue was purified by flash silica gel chromatography ( 20g The residue was purified by silica gel flash column (0-4% ethyl acetate / petroleum ether gradient @ 25 mL / min) to afford compound 2a (7.24 g, 53.1% yield) as a brown solid.

[0283] 1H NMR: (400MHz, CDCl3-d) δ7.42-7.56(m,1H),7.27-7.38(m,5H),7.03-7.15(m,1H),4.13(s,2H).

[0284] To a solution of compound 2a (4.50 g, 16.0 mmol, 1.00 eq.) in H 2 O (40.0 mL) and THF (8.00 mL) were added Zn(CN) 2 (2.16 g, 18.4 mmol, 1.17 mL, 1.15 eq.) and tBuXPhos Pd G 3 (1.27 g, 1.60 mmol, 0.100 eq.). The reaction mixture was stirred at 25° C. under N 2 atmosphere for 14 hours. TLC (petroleum ether:ethyl acetate=10:1, R f =0.06) indicated that compound 2a was completely consumed and a new spot was formed. The reaction was judged to be complete according to the TLC result. The reaction mixture was quenched with 30.0 mL of NaClO aqueous solution at 25 ° C, and then extracted with 90.0 mL (30.0 mL×3) of ethyl acetate. The combined organic layer was washed with 90.0 mL (30.0 mL×3) of brine, dried over Na2SO4, filtered and concentrated under reduced pressure to obtain a residue. The residue was purified by flash silica gel chromatography ( 40g The residue was purified by silica gel flash column (0-9% ethyl acetate / petroleum ether gradient @ 30 mL / min as eluent) to afford compound 3a as a brown solid (3.12 g, yield 77.2%, purity 90.0%).

[0285] LCMS: m / z=228.1 (M+H) + , Rt=2.400min.

[0286] 1 H NMR: (400MHz, CDCl3-d) δ7.69 (d, J=4.0Hz, 1H), 7.60 (d, J=4.0Hz, 1H), 7.24-7.43 (m, 5H), 4.22 (s, 2H).

[0287] To a solution of compound 3a (3.12 g, 13.7 mmol, 1.00 eq.) and formaldehyde (5.57 g, 68.6 mmol, 5.11 mL, 5 eq.) in MeOH (25.0 mL) were added piperidine (1.17 g, 13.7 mmol, 1.36 mL, 1.00 eq.) and acetic acid (1.65 g, 27.4 mmol, 1.57 mL, 2.00 eq.). The mixture was stirred at 70° C. for 2 hours. TLC (petroleum ether:ethyl acetate=10:1, Rf =0.32) indicated that compound 3a was completely consumed and a new spot was formed. The reaction was judged to be complete according to TLC results. The reaction mixture was concentrated under reduced pressure to obtain compound 4a (3.28 g, crude product) as a yellow oil.

[0288] LCMS: m / z=240.1 (M+H) + , Rt=2.549min.

[0289] Compound 4a (1.30 g, 5.43 mmol, 1.00 eq.) and hydrazine hydrate (810 mg, 16.2 mmol, 785 μL, 2.98 eq.) were dissolved in EtOH (10.0 mL). The mixture was stirred at 80° C. for 1 hour. LCMS (Rt = 1.092 min) showed complete consumption of compound 4a, with a major peak of the desired mass detected. The reaction mixture was concentrated under reduced pressure to afford compound 5a (1.40 g, crude) as a yellow solid.

[0290] m / z=254.0(M+H) + , Rt=1.092min.

[0291] Compound 5aa (1.10 g, 3.19 mmol, 1.00 eq.) was dissolved in toluene (2.00 mL). Compound 5a (807.01 mg, 3.19 mmol, 1.00 eq.) was then added. The mixture was stirred at 110°C for 2 hours. LCMS (Rt = 2.765 min) showed complete consumption of compound 5a, with a major peak of the desired mass detected. The reaction mixture was filtered and concentrated under reduced pressure to obtain a residue. The residue was purified by preparative HPLC (column: CD18-Welch Ultimate C18 150 × 40 × 7 μm; mobile phase: [H2O (0.05% HCl)-ACN]; gradient: 51% to 81% B over 15.0 min) to obtain compound 6a (430 mg, 21.4% yield, 80.0% purity) as a yellow solid.

[0292] LCMS: m / z=504.9 (M+H) + , Rt=2.765min.

[0293] LCMS: m / z=504.5 (M+H) + , Rt=2.873min.

[0294] 1H NMR: (400MHz, CDCl3-d)δ8.68(s,1H),8.32(d,J=8.6Hz,2H),7.85(d,J=8.0Hz,2H),7.33-7.45(m,5H ),7.16-7.18(m,1H),6.93(d,J=4.0Hz,1H),4.62-4.74(m,1H),4.35-4.49(m,1H),3.87-4.00(m,1H).

[0295] 19 F NMR: (376MHz, CDCl3-d)δ-63.239(s,3F).

[0296] Compound 6a (120 mg, 238 μmol, 1.00 eq.) was dissolved in toluene (5.00 mL). POCl (72.9 mg, 476 μmol, 44.3 μL, 2.00 eq.) and DIEA (61.5 mg, 476 μmol, 82.9 μL, 2.00 eq.) were then added. The mixture was stirred at 110° C. for 2 hours. LCMS (Rt=2.929 min) showed that compound 6a was completely consumed, and a major peak with the desired mass was detected. The reaction mixture was quenched with 1 M NaOH (20.0 mL) at 20° C. and then extracted with ethyl acetate (10.0 mL×3). The combined organic layers were washed with brine (10.0 mL), dried over NaSO, filtered, and concentrated under reduced pressure to obtain compound 7a (124 mg, crude product) as a brown oil.

[0297] LCMS: m / z=522.9 (M+H) + , Rt=2.929min.

[0298] Compound 7a (309 mg, 591 μmol, 1.00 eq.) was dissolved in DCM (5.00 mL). Compound 7 (122 mg, 886 μmol, 1.50 eq.) and TEA (299 mg, 2.95 mmol, 411 μL, 5.00 eq.) were then added. The mixture was stirred at 25° C. for 2 hours. TLC (petroleum ether:ethyl acetate=1:2, R f =0.43) showed that multiple new spots were formed. The reaction was judged to be complete according to the TLC results. The reaction mixture was filtered and concentrated under reduced pressure to obtain a residue. The residue was purified by flash silica gel chromatography ( 4g The product was purified by silica gel flash column (eluent: 0-75% ethyl acetate / petroleum ether gradient @ 17 mL / min), followed by preparative HPLC (column: CD24-WePure Biotech XPT C18 150×25×7 μm; mobile phase: [H2O (0.1% TFA)-ACN]; gradient: 37%-67% B in 10.0 min) to afford compound 8 (45.0 mg, 12.2% yield, 99.7% purity) as a white solid.

[0299] LCMS: m / z=668.0 (M+H) + , Rt=2.649min.

[0300] HPLC: Rt = 3.841 min.

[0301] 1H NMR: (400MHz, CDCl3-d) δ8.06 (d, J = 8.1Hz, 2H), 7.71 (br d, J = 6.8Hz, 2H), 7.53 (br s,1H),7.33-7.41(m,3H),7.14-7.24(m,3H),6.84-6.93(m,1H),4.72-4.81(m,2H),4.56-4 .71(m,2H),4.04-4.19(m,2H),3.90-4.02(m,1H),3.40-3.58(m,1H),1.49(d,J=7.0Hz,3H).

[0302] 19 F NMR: (376MHz, CDCl3-d)δ-62.958 (s, 3F).

[0303] Compound 8 was purified by SFC: chromatographic column: DAICEL CHIRALPAK AD (250 mm × 30 mm, 10 μm); mobile phase: [CO2-EtOH (0.1% NH3H2O)]; B%: 50%, isocratic elution mode, to obtain compound Q7-P1 (14.0 mg, yield 38.9%, purity 97.15%) as a white solid, and compound Q7-P2 (15.0 mg, yield 40.9%, purity 95.3%) as a white solid.

[0304] LCMS: m / z=624.5 (M+H) + , Rt=2.644min.

[0305] Compound Q7-P1:

[0306] LCMS: m / z=625.2 (M+H) +, Rt = 1.844 min.

[0307] HPLC: Rt = 3.842 min.

[0308] SFC: Rt = 0.457 min.

[0309] 1 H NMR: (400 MHz, CDCl3-d) δ 8.06 (d, J = 8.1 Hz, 2H), 7.72 (d, J = 8.3 Hz, 2H), 7.51 - 7.63 (m, 1H), 7.33 - 7.41 (m, 4H), 7.20 (dd, J = 7.8, 1.5 Hz, 2H), 6.93 (d, J = 4.0 Hz, 1H), 4.71 - 4.79 (m, 2H), 4.59 - 4.70 (m, 2H), 4.07 - 4.18 (m, 2H), 3.91 - 4.00 (m, 1H), 3.44 - 3.55 (m, 1H), 1.50 (d, J = 7.0 Hz, 3H).

[0310] 19 F NMR: (376 MHz, CDCl3-d) δ -62.951 (s, 3F).

[0311] Compound Q7-P2:

[0312] LCMS: m / z = 624.5 (M+H) + , Rt = 2.643 min.

[0313] HPLC: Rt = 3.839 min.

[0314] SFC: Rt = 1.210 min.

[0315] 1 H NMR: (400 MHz, CDCl3-d) δ 8.02 - 8.10 (m, 2H), 7.69 - 7.75 (m, 2H), 7.54 - 7.62 (m, 1H), 7.33 - 7.43 (m, 4H), 7.20 (dd, J = 7.8, 1.5 Hz, 2H), 6.93 (d, J = 4.0 Hz, 1H), 4.73 - 4.83 (m, 2H), 4.66 (d, J = 5.1 Hz, 2H), 4.04 - 4.22 (m, 2H), 3.87 - 4.02 (m, 1H), 3.43 - 3.58 (m, 1H), 1.50 (d, J = 7.0 Hz, 3H).

[0316] 19 F NMR: (376 MHz, CDCl3-d) δ -62.958 (s, 3F).

[0317] Preparation Example 25: Synthesis of (Z)-4-phenyl-N-(2-sulfamoylethyl)-N'-((4-(trifluoromethyl)phenyl)sulfonyl)-3-(5-(trifluoromethane)thiophen-2-yl)-4,5-dihydro-1H-pyrazole-1-carboximide (S, R enantiomers of Compound Q8)

[0318] To a solution of compound 1 (6.12 g, 72.8 mmol, 5.76 mL, 1.50 eq.) and compound 1a (7.50 g, 48.5 mmol, 6.47 mL, 1.00 eq.) in DCM (100.0 mL) was added AlCl₃ (9.70 g, 72.8 mmol, 3.98 mL, 1.50 eq.) at 0°C. The reaction mixture was stirred at 25°C under a N₂ atmosphere for 1 hour. TLC (petroleum ether:ethyl acetate = 10:1, R f =0.60) indicated that compound 1 was completely consumed and two new spots were formed. The reaction mixture was quenched with 1.00M HCl (60.0 mL) and extracted with DCM (500.0 mL×3). The combined organic layers were washed with brine (600.0 mL×2), dried over Na2SO4, filtered and concentrated under reduced pressure to give a residue. The residue was purified by flash silica gel chromatography ( 20g The product was purified by silica gel flash column (0-4% ethyl acetate / petroleum ether gradient @ 25 mL / min) to afford compound 2 as a brown oil (8.10 g, 82.54% yield).

[0319] 1 H NMR: (400MHz, CDCl3-d) δ7.70 (d, J = 3.5Hz, 1H), 7.56 (d, J = 4.9Hz, 1H), 7.20-7.33 (m, 4H), 7.13-7.20 (m, 1H), 7.05 (t, J = 4.4Hz, 1H), 4.12 (s, 2H).

[0320] A 40.0 mL vial containing a stir bar was charged with a solution of compound 2 (8.00 g, 39.6 mmol, 1.00 eq.), Ru(bpy)3Cl2·6H2O (237 mg, 316 μmol, 0.00800 eq.), and 1-oxo-4-phenylpyridin-1-ium (13.5 g, 79.1 mmol, 2.00 eq.). The vial was sealed and placed under nitrogen. (2,2,2-trifluoroacetyl) 2,2,2-trifluoroacetate (20.8 g, 98.9 mmol, 13.7 mL, 2.50 eq.) was then added. The reaction was stirred and illuminated with a 4×50 W blue LED lamp (3 cm away). The reaction temperature was maintained at 25°C with cooling water for 14 hours. TLC (petroleum ether:ethyl acetate = 0:1, R f =0.27) indicated that compound 2 had reacted completely and formed multiple new spots. The reaction was judged to be complete according to TLC results. The reaction mixture was filtered and concentrated under reduced pressure to obtain a residue. The residue was purified by flash silica gel chromatography ( 80g The product was purified by silica gel flash column with 0% ethyl acetate / petroleum ether gradient @ 50 mL / min as eluent to afford compound 3 (8.00 g, 74.8% yield) as a brown oil.

[0321] To a solution of compound 3 (6.25 g, 23.1 mmol, 1.00 eq.) and formaldehyde (9.38 g, 116 mmol, 8.61 mL, 5.00 eq.) in MeOH (50.0 mL) were added piperidine (1.97 g, 23.1 mmol, 2.28 mL, 1.00 eq.) and acetic acid (2.78 g, 46.2 mmol, 2.65 mL, 2.00 eq.). The mixture was stirred at 70° C. for 2 hours. LCMS (Rt=2.116 min) indicated complete consumption of compound 3, with a major peak of the desired mass detected. The reaction mixture was filtered and concentrated under reduced pressure to obtain a residue. The residue was purified by preparative HPLC (chromatographic column: CD18-Welch Ultimate C18 150×40×7 μm; mobile phase: [H2O (0.1% TFA)-ACN]; gradient: 53%-83% B in 15.0 min) to give compound 4 (1.079 g, yield 14.9%, purity 90%) as a brown oil.

[0322] LCMS: m / z=283.2 (M+H) + , Rt=2.116min.

[0323] LCMS: m / z=282.8 (M+H) +, Rt=3.018min.

[0324] 1 H NMR: (400MHz, CDCl3-d) δ7.50 (br d, J = 2.6Hz, 1H), 7.41-7.45 (m, 2H), 7.35-7.41 (m, 4H), 5.84-6.11 (m, 2H).

[0325] Compound 4 (1.08 g, 3.82 mmol, 1.00 eq.) and hydrazine hydrate (900 mg, 18.0 mmol, 872 μL, 4.70 eq.) were dissolved in EtOH (10.0 mL). The mixture was stirred at 80 ° C for 1 hour. LCMS (Rt = 2.722 min) showed that compound 4 was completely consumed and a main peak with the desired mass number was detected. The reaction mixture was concentrated under reduced pressure to obtain a residue. EtOH (20.0 mL) was added to the residue at 0 ° C, and a yellow solid precipitate was observed, which was then filtered through a sand core funnel and washed with EtOH (5.00 mL × 3). The yellow filter cake was concentrated under reduced pressure to obtain a residue. The residue was then purified by preparative HPLC (chromatographic column: CD43-Welch Ultimate XB-CN 250×50×10 μm; mobile phase: [hexane-EtOH]; gradient: 1%-10% B in 15.0 min) to give white solid compound 5 (1.00 g, yield 88.3%).

[0326] LCMS: m / z=296.8 (M+H) + , Rt=2.722min.

[0327] 1 H NMR: (400MHz, CDCl3-d)δ7.29-7.40(m,5H),7.12-7.16(m,1H),6.64(dd,J=3.8,1.1Hz,1H ), 4.46 (dd, J = 10.8, 7.3Hz, 1H), 4.07 (dd, J = 10.8, 9.6Hz, 1H), 3.59 (dd, J = 9.5, 7.3Hz, 1H).

[0328] 19 F NMR: (376MHz, CDCl3-d)δ-55.685(s,3F).

[0329] Compound 5a (1.70 g, 4.92 mmol, 1.00 eq.) was dissolved in toluene (5 mL). Compound 5 (1.46 g, 4.92 mmol, 1.00 eq.) was then added. The mixture was stirred at 110°C for 1 hour. LCMS (Rt = 3.106 min) showed complete consumption of compound 5, with a major peak of the desired mass detected. The reaction mixture was filtered and concentrated under reduced pressure to obtain a residue. The residue was purified by preparative HPLC (column: CD18-Welch Ultimate C18 150 × 40 × 7 μm; mobile phase: [H2O (0.1% TFA)-ACN]; gradient: 51% to 81% B over 15.0 min) to obtain compound 6 (1.70 g, 56.8% yield, 90.0% purity) as a yellow solid.

[0330] LCMS: m / z=547.5 (M+H) + , Rt=3.106min.

[0331] LCMS: m / z=547.5 (M+H) + , Rt=3.054min.

[0332] 1 H NMR: (400MHz, CDCl3-d)δ8.62-8.75(m,1H),8.31(d,J=8.3Hz,2H),7.79-7.90(m,2H),7.33-7.40(m, 4H),7.17-7.21(m,2H),6.83-6.92(m,1H),4.66-4.75(m,1H),4.28-4.48(m,1H),3.83-4.01(m,1H).

[0333] Compound 6 (213 mg, 389 μmol, 1.00 eq.) was dissolved in toluene (2.00 mL). POCl₃ (119 mg, 778 μmol, 72.5 μL, 2.00 eq.) and DIEA (100 mg, 778 μmol, 136 μL, 2.00 eq.) were then added. The mixture was stirred at 110°C for 1 hour. LCMS (Rt = 2.378 min) showed complete consumption of compound 6, with a major peak of the desired mass detected. The reaction mixture was quenched with 1 M NaOH (20.0 mL) at 20°C and then extracted with ethyl acetate (10.0 mL x 3). The combined organic layers were washed with brine (10.0 mL), dried over Na₂SO₄, filtered, and concentrated under reduced pressure to afford compound 7 (220 mg, crude) as a brown oil.

[0334] LCMS: m / z=566.1 (M+H) + , Rt=2.378min.

[0335] Compound 7 (220 mg, 389 μmol, 1.00 eq.) was dissolved in DCM (3.00 mL). TEA (197 mg, 1.94 mmol, 270 μL, 5.00 eq.) and compound 7a (93.7 mg, 583 μmol, 1.50 eq.) were then added. The mixture was stirred at 25° C. for 2 hours. TLC (petroleum ether:ethyl acetate=1:2, R f =0.41) indicating that compound 7 was completely consumed and many new spots were formed. The reaction was determined to be complete based on TLC results. The reaction mixture was filtered and concentrated under reduced pressure to obtain a residue. The residue was purified by preparative TLC (SiO2, petroleum ether / ethyl acetate = 1 / 2) to obtain compound 8 (180 mg, 70.7% yield, 99.7% purity) as a white solid.

[0336] LCMS: m / z=654.1 (M+H) + , Rt=2.070min.

[0337] HPLC: Rt=3.143 min.

[0338] SFC: Rt=1.555, 1.743min.

[0339] 1H NMR: (400MHz, CDCl3-d) δ8.01-8.10 (m, 2H), 7.71 (d, J = 8.3Hz, 2H), 7.45-7.54 (m,1H),7.30-7.42(m,3H),7.15-7.24(m,3H),6.88(d,J=3.8Hz,1H),4.94(br s, 2H), 4.57-4.72 (m, 2H), 4.07-4.25 (m, 3H), 3.53 (t, J = 5.9Hz, 2H).

[0340] 19 F NMR: (376MHz, CDCl3-d)δ-55.938(s,3F),-62.947(s,3F).

[0341] Compound 8 was purified by SFC: chromatographic column: DAICEL CHIRALCEL OD (250 mm × 30 mm, 10 μm); mobile phase: [CO2-MeOH (0.1% NH3H2O)]; B%: 45%, isocratic elution mode to obtain compound Q8-P1 (65.0 mg, yield 36.1%, purity 99.9%) as a white solid, and compound Q8-P2 (65.0 mg, yield 36.0%, purity 99.6%) as a white solid.

[0342] LCMS: m / z=653.4 (M+H) + , Rt=2.851min.

[0343] Compound Q8-P1:

[0344] LCMS: m / z=654.1 (M+H) + , Rt=1.574min.

[0345] HPLC Rt = 4.156 min.

[0346] SFC: Rt = 1.892 min.

[0347] 1 H NMR: (400MHz, CDCl3-d)δ8.05(d,J=8.3Hz,2H),7.72(d,J=8.3Hz,2H),7.43-7.52(m,1H),7.32-7.41(m,3H),7.1 7-7.24(m,3H),6.88(d,J=3.8Hz,1H),4.91(s,2H),4.56-4.72(m,2H),4.08-4.24(m,3H),3.52(t,J=5.9Hz,2H).

[0348] 19 F NMR: (377MHz, CDCl3-d)δ-55.941(s,3F),-62.951(s,3F).

[0349] Compound Q8-P2:

[0350] LCMS: m / z=654.2 (M+H) + , Rt=1.575min.

[0351] HPLC: Rt = 4.153 min.

[0352] SFC: Rt = 2.043 min.

[0353] 1H NMR: (400MHz, CDCl3-d)δ8.05(d,J=8.4Hz,2H),7.71(d,J=8.4Hz,2H),7.44-7.51(m,1H),7.31-7.42(m,3H),7 .15-7.23(m,3H),6.88(d,J=3.0Hz,1H),4.93(s,2H),4.51-4.77(m,2H),4.03-4.29(m,3H),3.41-3.63(m,2H).

[0354] 19 F NMR: (376MHz, CDCl3-d)δ-55.948(s,3F),-62.951(s,3F).

[0355] Preparation Example 26: Synthesis of (Z)-4-phenyl-N-((R)-2-sulfamoylpropyl)-N'-((4-(trifluoromethyl)phenyl)sulfonyl)-3-(5-(trifluoromethane)thiophen-2-yl)-4,5-dihydro-1H-pyrazole-1-carboximide (S, R enantiomers of compound Q9)

[0356] To a solution of compound 1 (25.0 g, 333 mmol, 26.2 mL, 1.00 eq.) in toluene (250 mL) was added Et3N (3.37 g, 33.3 mmol, 4.63 mL, 0.100 eq.) and compound 1a (49.3 g, 333 mmol, 1.00 eq.). The mixture was stirred at 130 ° C for 3 hours. LC-MS (Rt = 2.334 min) showed that compound 1 was completely consumed and a peak of the desired mass was detected. The reaction mixture was filtered and concentrated under reduced pressure to give compound 2 (68.3 g, crude product) as a white solid.

[0357] LCMS: m / z=206.1 (M+H) + , Rt=2.334min.

[0358] To a solution of compound 2 (68.3 g, 333 mmol, 1.00 eq.) in DCM (250 mL) at 0°C were added TEA (101 g, 998 mmol, 139 mL, 3.00 eq.) and DMAP (407 mg, 3.33 mmol, 0.0100 eq.), followed by 4-methylbenzenesulfonyl chloride (69.8 g, 366 mmol, 1.10 eq.). The mixture was stirred at 25°C for 10 hours. LC-MS (Rt = 2.331 min) showed that compound 2 was completely consumed, and a peak of the desired mass was detected. The reaction mixture was filtered and concentrated under reduced pressure to obtain a residue. The crude product was triturated with 50.0 mL of (PE:EA = 10:1) at 25°C for 30 min to obtain compound 3 (89.0 g, 74.4% yield) as a white solid.

[0359] LCMS: m / z=360.1 (M+H) + , Rt=2.331min.

[0360] Compound 3 (38.1 g, 106 mmol, 1.00 eq.) and compound 3a (13.1 g, 117 mmol, 1.10 eq.) were added to DMF (200 mL), followed by anhydrous KCO (29.3 g, 212 mmol, 2.00 eq.), and the mixture was stirred at 60°C for 5 hours. LC-MS (1.618 min) indicated the detection of the desired compound. The reaction mixture was extracted with 180 mL of EA (60.0 mL x 3). The combined organic layers were washed with 180 mL of water (60.0 mL x 3), dried over NaSO, filtered, and concentrated under reduced pressure to yield a residue. The residue was purified by preparative HPLC (chromatographic column: Ultimate XB-SIO2 250×50 mm×10 μm; mobile phase: [hexane-EtOH]; gradient: 1%-26% B in 15.0 min) to give white solid compound 4 (20.0 g, yield 63.0%).

[0361] LCMS: m / z=299.8 (M+H) + , Rt=1.618min.

[0362] Compound 4 (8.30 g, 27.7 mmol, 1.00 eq.) was dissolved in DCM (15.0 mL). mCPBA (8.44 g, 41.6 mmol, 1.50 eq.) was then added at 0°C. The mixture was stirred at 25°C for 10 hours. TLC (petroleum ether:ethyl acetate = 1:1, R f=0.08) indicated that compound 4 was completely consumed and multiple new spots appeared. The reaction mixture was quenched with saturated aqueous Na2S2O3 solution (10 mL) and extracted with EtOAc (10.0 mL×2). The combined organic layers were washed with saturated aqueous NaHCO3 solution (10.0 mL) and brine (10.0 mL), dried over Na2SO4, filtered and concentrated under reduced pressure. The residue was purified by flash silica gel chromatography ( 40g The product was purified by silica gel flash column (eluting with 0-55% ethyl acetate / petroleum ether gradient @ 40 mL / min) to afford compound 5 (8.89 g, 96.7% yield) as a white solid.

[0363] 1 H NMR: (400MHz, CDCl3-d) δ8.95 (d, J = 4.9Hz, 2H), 7.79-7.88 (m, 2H), 7.70-7.77 (m, 2H) ,7.55(t,J=4.8Hz,1H),4.23-4.40(m,2H),3.97-4.10(m,1H),1.51(d,J=7.0Hz,3H).

[0364] A solution of compound 5 (8.00 g, 24.1 mmol, 1.00 eq.) dissolved in MeOH (40.0 mL) was cooled to 0° C. and treated with sodium methoxide (5.00 M, 5.79 mL, 1.20 eq.). The mixture was stirred at this temperature for 2 hours and then treated with a solution of solid sodium acetate (2.97 g, 36.2 mmol, 1.50 eq.) and hydroxylamine sulfonic acid (4.10 g, 36.2 mmol, 1.50 eq.) dissolved in H O (40 mL), heated to 20° C. and maintained for 12 hours. TLC (petroleum ether: ethyl acetate = 1:1, R f =0.33) indicated that compound 5 was completely consumed and a new spot was formed. The reaction mixture was filtered and concentrated under reduced pressure to obtain a residue. The residue was purified by flash silica gel chromatography ( 40g The product was purified by silica gel flash column with a gradient of 0-50% ethyl acetate / petroleum ether at 45 mL / min as the eluent to afford compound 6 (2.63 g, 9.79 mmol, 40.5% yield) as a white solid.

[0365] 1H NMR: (400MHz, DMSO-d6) δ7.77-8.06 (m, 4H), 7.01 (s, 2H), 4.02 (dd, J = 14.0, 5.1 Hz, 1H), 3.72 (dd, J = 13.9, 9.7Hz, 1H), 3.37-3.42 (m, 1H), 1.21 (d, J = 6.9Hz, 3H).

[0366] Compound 6 (1.40 g, 5.22 mmol, 1.00 eq.) and NH2NH2·H2O (900 mg, 18.0 mmol, 872 μL, 100% purity, 3.45 eq.) were dissolved in EtOH (15.0 mL). The mixture was stirred at 80°C under N2 for 1 hour. LCMS (Rt = 0.410 min) showed complete consumption of compound 6, with a major peak of the desired mass detected. The reaction mixture was concentrated under reduced pressure to obtain a residue. The residue was purified by preparative HPLC (column: CD01-Phenomenex Luna C18 150 × 25 mm × 10 μm; mobile phase: [H2O (0.225% FA)-ACN]; gradient: 0% to 30% B over 10.0 min) to obtain compound 7 (1.20 g, 99.8% yield) as a colorless oil.

[0367] LCMS: m / z=139.1 (M+H) + , Rt=0.410min.

[0368] To a solution of compound 1a (6.12 g, 72.8 mmol, 5.76 mL, 1.50 eq.) and compound 1aa (7.50 g, 48.5 mmol, 6.47 mL, 1.00 eq.) in DCM (100.0 mL) was added AlCl₃ (9.70 g, 72.8 mmol, 3.98 mL, 1.50 eq.) at 0°C. The reaction mixture was stirred at 25°C under a N₂ atmosphere for 1 hour. TLC (petroleum ether:ethyl acetate = 10:1, R f =0.60) showed that compound 1a was completely consumed and two new spots were formed. The reaction mixture was quenched with 1.00M HCl (60.0 mL) and extracted with DCM (500.0 mL×3). The combined organic layers were washed with brine (600.0 mL×2), dried over Na2SO4, filtered and concentrated under reduced pressure to give a residue. The residue was purified by flash silica gel chromatography ( 20g The product was purified by silica gel flash column with a gradient of 0-4% ethyl acetate / petroleum ether at 25 mL / min as eluent to afford compound 2a (8.10 g, 82.54% yield) as a brown oil.

[0369] 1 H NMR: (400MHz, CDCl3-d) δ7.70 (d, J = 3.5Hz, 1H), 7.56 (d, J = 4.9Hz, 1H), 7.20-7.33 (m, 4H), 7.13-7.20 (m, 1H), 7.05 (t, J = 4.4Hz, 1H), 4.12 (s, 2H).

[0370] To a 40.0 mL reaction vial equipped with a stir bar was added a solution of compound 2a (8.00 g, 39.6 mmol, 1.00 eq.), Ru(bpy)3Cl2·6H2O (237 mg, 316 μmol, 0.00800 eq.), and 1-oxo-4-phenylpyridin-1-ium (13.5 g, 79.1 mmol, 2.00 eq.). The vial was sealed and placed under nitrogen. (2,2,2-trifluoroacetyl) 2,2,2-trifluoroacetate (20.8 g, 98.9 mmol, 13.7 mL, 2.50 eq.) was then added. The reaction was stirred and illuminated with a 4×50 W blue LED lamp (3 cm away). The reaction temperature was maintained at 25°C with cooling water for 14 hours. TLC (petroleum ether:ethyl acetate = 10:1, R f =0.27) indicated that compound 2a reacted completely and formed multiple new spots. The reaction was judged to be complete according to TLC results. The reaction mixture was filtered and concentrated under reduced pressure to obtain a residue. The residue was purified by flash silica gel chromatography ( 80g Purification by silica gel flash column, eluting with 0% ethyl acetate / petroleum ether gradient @ 50 mL / min) afforded compound 3a (8.00 g, 74.8% yield) as a brown oil.

[0371] To a solution of compound 3a (6.25 g, 23.1 mmol, 1.00 eq.) and formaldehyde (9.38 g, 116 mmol, 8.61 mL, 5.00 eq.) in MeOH (50.0 mL) were added piperidine (1.97 g, 23.1 mmol, 2.28 mL, 1.00 eq.) and acetic acid (2.78 g, 46.2 mmol, 2.65 mL, 2.00 eq.). The mixture was stirred at 70°C for 2 hours. LCMS (Rt = 2.116 min) indicated complete consumption of compound 3a, with a major peak of the desired mass detected. The reaction mixture was filtered and concentrated under reduced pressure to obtain a residue. The residue was purified by preparative HPLC (column: CD18-Welch Ultimate C18 150×40×7 μm; mobile phase: [H2O (0.1% TFA)-ACN]; gradient: 53%-83% B in 15.0 min) to give compound 4a (1.079 g, yield 14.9%, purity 90%) as a brown oil.

[0372] LCMS: m / z=283.2 (M+H) + , Rt=2.116min.

[0373] LCMS: m / z=282.8 (M+H) + , Rt=3.018min.

[0374] 1 H NMR: (400MHz, CDCl3-d) δ7.50 (br d, J = 2.6Hz, 1H), 7.41-7.45 (m, 2H), 7.35-7.41 (m, 4H), 5.84-6.11 (m, 2H).

[0375] Compound 4a (1.08 g, 3.82 mmol, 1.00 eq.) and hydrazine hydrate (900 mg, 18.0 mmol, 872 μL, 4.70 eq.) were dissolved in EtOH (10.0 mL). The mixture was stirred at 80° C. for 1 hour. LCMS (Rt=2.722 min) showed that compound 4a was completely consumed, and a main peak with the desired mass number was detected. The reaction mixture was concentrated under reduced pressure to obtain a residue. EtOH (20.0 mL) was added to the residue at 0° C., and a yellow solid precipitate was observed, which was then filtered through a sand core funnel and washed with EtOH (5.00 mL×3). The yellow filter cake was concentrated under reduced pressure to obtain a residue. The residue was then purified by preparative HPLC (column: CD43-Welch Ultimate XB-CN 250×50×10 μm; mobile phase: [hexane-EtOH]; gradient: 1%-10% B in 15.0 min) to give white solid compound 5a (1.00 g, yield 88.3%).

[0376] LCMS: m / z=296.8 (M+H) + , Rt=2.722min.

[0377] 1 H NMR: (400MHz, CDCl3-d)δ7.29-7.40(m,5H),7.12-7.16(m,1H),6.64(dd,J=3.8,1.1Hz,1H ), 4.46 (dd, J = 10.8, 7.3Hz, 1H), 4.07 (dd, J = 10.8, 9.6Hz, 1H), 3.59 (dd, J = 9.5, 7.3Hz, 1H).

[0378] 19 F NMR: (376MHz, CDCl3-d)δ-55.685(s,3F).

[0379] Compound 5aa (1.70 g, 4.92 mmol, 1.00 eq.) was dissolved in toluene (5 mL). Compound 5a (1.46 g, 4.92 mmol, 1.00 eq.) was then added. The mixture was stirred at 110°C for 1 hour. LCMS (Rt = 3.106 min) showed complete consumption of compound 5aa, with a major peak of the desired mass detected. The reaction mixture was filtered and concentrated under reduced pressure to obtain a residue. The residue was purified by preparative HPLC (column: CD18-Welch Ultimate C18 150 × 40 × 7 μm; mobile phase: [H2O (0.1% TFA)-ACN]; gradient: 51% to 81% B over 15.0 min) to obtain compound 6a (1.70 g, 56.8% yield, 90.0% purity) as a yellow solid.

[0380] LCMS: m / z=547.5 (M+H) + , Rt=3.106min.

[0381] LCMS: m / z=547.5 (M+H) + , Rt=3.054min.

[0382] 1 H NMR: (400MHz, CDCl3-d)δ8.62-8.75(m,1H),8.31(d,J=8.3Hz,2H),7.79-7.90(m,2H),7.33-7.40(m, 4H),7.17-7.21(m,2H),6.83-6.92(m,1H),4.66-4.75(m,1H),4.28-4.48(m,1H),3.83-4.01(m,1H).

[0383] Compound 6a (213 mg, 389 μmol, 1.00 eq.) was dissolved in toluene (2.00 mL). POCl (119 mg, 778 μmol, 72.5 μL, 2.00 eq.) and DIEA (100 mg, 778 μmol, 136 μL, 2.00 eq.) were then added. The mixture was stirred at 110° C. for 1 hour. LCMS (Rt=2.378 min) showed that compound 6a was completely consumed, and a major peak with the desired mass was detected. The reaction mixture was quenched with 1 M NaOH (20.0 mL) at 20° C. and then extracted with ethyl acetate (10.0 mL×3). The combined organic layers were washed with brine (10.0 mL), dried over NaSO, filtered, and concentrated under reduced pressure to obtain compound 7a (220 mg, crude product) as a brown oil.

[0384] LCMS: m / z=566.1 (M+H) + , Rt=2.378min.

[0385] 1 H NMR: (400MHz, DMSO-d6) δ3.03-3.10 (m, 2H), 2.69-2.81 (m, 1H), 1.26 (d, J = 6.8Hz, 3H).

[0386] Compound 7a (200 mg, 353 μmol, 1.00 eq.) was dissolved in DCM (4.00 mL). Compound 7 (97 mg, 707 μmol, 2.00 eq.) and TEA (179 mg, 1.77 mmol, 246 μL, 5.00 eq.) were then added. The mixture was stirred at 25°C for 2 hours. LCMS (Rt = 2.855 min) showed complete consumption of compound 7a, with a major peak of the desired mass detected. The reaction mixture was filtered and concentrated under reduced pressure to obtain a residue. The residue was purified by preparative HPLC (column: CD24-WePure Biotech XPT C18 150 × 25 × 7 μm; mobile phase: [H2O (0.1% TFA)-ACN]; gradient: 43% to 73% B over 10.0 min) to obtain compound 8 (50.0 mg, 19.8% yield, 93.3% purity) as a white solid.

[0387] LCMS: m / z=667.4 (M+H) + , Rt=2.855min.

[0388] LCMS: m / z=668.0 (M+H) + , Rt=2.875min.

[0389] HPLC: EC26335-198-P1H3, Rt=4.237min.

[0390] 1H NMR: (400MHz, CDCl3-d) δ8.06 (d, J = 8.1Hz, 2H), 7.71 (br d, J = 6.8Hz, 2H), 7.53 (br s,1H),7.33-7.41(m,3H),7.14-7.24(m,3H),6.84-6.93(m,1H),4.72-4.81(m,2H),4.56-4 .71(m,2H),4.04-4.19(m,2H),3.90-4.02(m,1H),3.40-3.58(m,1H),1.49(d,J=7.0Hz,3H).

[0391] 19 F NMR: (376MHz, CDCl3-d)δ-55.933(s,3F),-62.943(s,3F).

[0392] Compound 8 was purified by SFC: chromatographic column: DAICEL CHIRALPAK AD (250 mm × 30 mm, 10 μm); mobile phase: [CO2-IPA (0.1% NH3H2O)]; B%: 42%, isocratic elution mode, to obtain compound Q9-P1 (10.0 mg, yield 19.5%, purity 97.5%) as a white solid, and compound Q9-P2 (13.5 mg, yield 26.6%, purity 98.7%) as a white solid.

[0393] LCMS: m / z=667.4 (M+H) + , Rt=2.849min

[0394] Compound Q9-P1:

[0395] LCMS: m / z=668.1 (M+H) + , Rt=1.593min.

[0396] HPLC: Rt = 4.242 min.

[0397] SFC: Rt = 1.722 min.

[0398] 1 H NMR: (400MHz, CDCl3-d) δ7.98-8.12(m,2H),7.66-7.78(m,2H),7.47-7.62(m,1H),7.31-7.42(m,3H),7.13-7.22(m,3H),6.85-6. 93(m,1H),4.76-4.97(m,2H),4.54-4.73(m,2H),4.03-4.22(m,2H),3.88-4.01(m,1H),3.42-3.57(m,1H),1.49(d,J=6.9Hz,3H).

[0399] 19 F NMR: (376MHz, CDCl3-d)δ-55.933(s,3F),-62.943(s,3F).

[0400] Compound Q9-P2:

[0401] LCMS: m / z=668.2 (M+H) +, Rt=2.100min.

[0402] HPLC: Rt = 4.237 min.

[0403] SFC: Rt = 1.946 min.

[0404] 1 H NMR: (400MHz, CDCl3-d)δ8.06(d,J=8.8Hz,2H),7.71(d,J=8.4Hz,2H),7.50-7.55(m,1H),7.34-7.41(m,3H),7.19-7.22(m,3H),6.86 -6.90(m,1H),4.75(s,2H),4.60-4.68(m,2H),4.04-4.14(m,2H),3.92(s,1H),3.50(quind,J=7.1,4.3Hz,1H),1.50(d,J=6.9Hz,3H).

[0405] 19 F NMR: (376MHz, CDCl3-d)δ-55.933(s,3F),-62.943(s,3F).

[0406] Preparation Example 27: Synthesis of (Z)-3-(2-cyanothiazol-5-yl)-4-phenyl-N-(2-sulfamoylethyl)-N'-((4-(trifluoromethyl)phenyl)sulfonyl)-4,5-dihydro-1H-pyrazole-1-carboximide (S, R enantiomers of Compound Q10)

[0407] To a solution of compound 1 (25.0 g, 152 mmol) in DCM (250 mL) were added EDCI (43.9 g, 229 mmol) and TEA (49.4 g, 489 mmol, 68.0 mL). Compound 1a (19.3 g, 198 mmol) was then added. The mixture was stirred at 25 ° C for 12 hours. TLC (petroleum ether: ethyl acetate = 5:1, R f =0.15) indicated that compound 1 was completely consumed and a new spot was formed. The reaction mixture was filtered and concentrated under reduced pressure to obtain a residue. The residue was purified by flash silica gel chromatography ( 40g The product was purified by silica gel flash column with a gradient of 0-15% ethyl acetate / petroleum ether at 45 mL / min as the eluent to afford compound 2 (30.2 g, 146 mmol, 86.1% yield, 90.0% purity) as a white solid.

[0408] 1H NMR: (400MHz, CDCl3-d) δ8.32(s,1H),3.79(s,3H),3.37(s,3H).

[0409] To a solution of compound 2 (17.5 g, 84.6 mmol) in THF (180 mL) was added dropwise compound 2a (1 M, 127 mL) at 0°C under N2 atmosphere. The mixture was stirred at 0°C for 3 hours. TLC (petroleum ether:ethyl acetate = 5:1, R f =0.40) indicated that compound 2 was completely consumed and a new spot was formed. The reaction mixture was quenched by adding saturated NH4Cl aqueous solution (300 mL) at 0°C and N2 atmosphere, and then extracted with ethyl acetate (300 mL×2). The combined organic layers were washed with brine (300 mL), dried over Na2SO4, filtered and concentrated under reduced pressure to obtain a residue. The residue was purified by flash silica gel chromatography ( 200g The product was purified by silica gel flash column with a gradient of 0-15% ethyl acetate / petroleum ether at 40 mL / min as eluent to afford compound 3 as a yellow oil (12.5 g, 47.3 mmol, 55.8% yield, 90.0% purity).

[0410] 1 H NMR: (400MHz, CDCl3-d) δ8.10 (s, 1H), 7.25-7.38 (m, 5H), 4.14 (s, 2H).

[0411] To a solution of compound 3 (6.00 g, 25.2 mmol) and formaldehyde (10.2 g, 126 mmol, 9.40 mL) in MeOH (60.0 mL) were added piperidine (2.15 g, 25.2 mol, 2.49 mL) and AcOH (3.03 g, 50.4 mmol, 2.89 mL). The mixture was stirred at 50° C. for 1 hour. LCMS (Rt=2.604 minutes) showed that compound 3 was completely consumed, and a major peak with the desired mass was detected. The reaction mixture was filtered and concentrated under reduced pressure to give compound 4 (6.30 g, crude product) as a yellow oil.

[0412] LCMS: m / z=250.1 (M+H) + ,Rt=2.604min.

[0413] To a solution of N2H4.H2O (1.36 g, 27.1 mmol, 1.32 mL) in ethanol (60.0 mL) was added a solution of compound 4 (6.00 g, 24.0 mmol) in ethanol (60.0 mL) dropwise over 10 minutes. The reaction mixture was stirred at 25°C for 2 hours. LCMS (Rt = 2.309 minutes) showed complete consumption of compound 4, and a major peak with the desired mass was detected. The reaction mixture was concentrated under reduced pressure to afford INT_7 (6.34 g, crude) as a yellow solid.

[0414] LCMS: m / z=264.1 (M+H) + ,Rt=2.309min.

[0415] To a solution of compound 5 (7.50 g, 21.7 mmol) in toluene (70.0 mL) was added INT_7 (5.73 g, 21.7 mmol). The mixture was stirred at 110 ° C for 1 hour. LCMS (Rt = 2.807 minutes) showed that INT_7 was completely consumed and a main peak with the desired mass was detected. The reaction mixture was filtered and concentrated under reduced pressure to obtain a residue. The residue was purified by preparative HPLC (TFA conditions) (chromatographic column: CD34-YMC Triart C18 250×70 mm×7 μm; mobile phase: [H2O (0.01% TFA)-ACN]; gradient: 40%-70% B in 30.0 min) to give a yellow oily compound 6 (1.80 g, 1.98 mmol, 9.12% yield, 56.6% purity).

[0416] LCMS: m / z=515.0 (M+H) + ,Rt=2.807min.

[0417] LCMS: m / z=515.0 (M+H) + ,Rt=2.808min.

[0418] A solution of NaCN (343 mg, 7.00 mmol) in H2O (1.80 mL) was slowly added to a stirred solution of compound 6 (1.80 g, 3.50 mmol) and DABCO (98.0 mg, 873 μmol, 96.1 μL) in DMSO (18.0 mL). Under nitrogen, the resulting solution was stirred at 50 ° C for 2 hours. LCMS (Rt = 2.744 minutes) showed that compound 6 was completely consumed, and a main peak with the required mass was detected. The reaction mixture was quenched to pH>11 by adding 2M NaOH at 0 ° C, and then saturated NaClO aqueous solution (20.0 mL) was added at 25 ° C to quench the reaction mixture, followed by extraction with ethyl acetate (20.0 mL × 3). The combined organic layers were washed with brine (20.0 mL), dried over Na2SO4, filtered and concentrated under reduced pressure to give compound 7 (1.10 g, crude product) as a yellow oil.

[0419] LCMS: m / z=506.0 (M+H) + ,Rt=2.744min.

[0420] To a solution of compound 7 (1.10 g, 2.18 mmol) in toluene (5.00 mL) was added POCl (667 mg, 4.35 mmol, 405 μL) and DIEA (562 mg, 4.35 mmol, 758 μL). The mixture was stirred at 110° C. for 1 hour. LCMS (Rt=2.897 minutes) showed that compound 7 was completely consumed, and a main peak with the desired mass was detected. 1 M NaOH (30.0 mL) was added to quench the reaction mixture at 0° C., followed by extraction with ethyl acetate (30.0 mL×3). The combined organic layers were washed with brine (30.0 mL), dried over Na SO , filtered, and concentrated under reduced pressure to give compound 8 (1.05 g, crude product) as a yellow oil.

[0421] LCMS: m / z=524.0 (M+H) + ,Rt=2.897min.

[0422] To a solution of compound 8 (1.05 g, 2.00 mmol) in DCM (10.5 mL) was added TEA (1.01 g, 10.0 mmol, 1.39 mL) and compound 8a (482 mg, 3.01 mmol). The mixture was stirred at 25 ° C for 2 hours. LCMS (Rt = 2.543 minutes) showed that compound 8 was completely consumed and a major peak with the desired mass was detected. The reaction mixture was filtered and concentrated under reduced pressure to obtain a residue. The residue was purified by flash silica gel chromatography ( 20g Purification by silica gel flash column, eluent: 0-50% petroleum ether:ethyl acetate, gradient @50 mL / min) afforded compound 9 (125 mg, 200 μmol, 9.98% yield, 97.88% purity) as a yellow solid.

[0423] LCMS: m / z=612.1 (M+H) + ,Rt=2.543min.

[0424] Compound 9:

[0425] LCMS: m / z=612.0 (M+H) + ,Rt=2.542min.

[0426] 1 H NMR: (400MHz, CDCl3-d)δ8.04(d,J=8.1Hz,2H),7.66-7.76(m,3H),7.49-7.58(m,1H),7.33-7.42(m ,3H),7.17-7.22(m,2H),5.02(s,2H),4.62-4.72(m,2H),4.13-4.22(m,3H),3.51(t,J=5.7Hz,2H).

[0427] Compound 9 (105 mg) was purified by SFC (chromatographic column: SFC-AD-30-DAICEL CHIRALPAK AD (250 mm×30 mm, 10 μm); mobile phase: [CO2-EtOH (0.1% NH3H2O)]; B%: 45%, isocratic elution mode) to obtain compound Q10-P1 (27.45 mg, purity 93.46%) as a yellow solid, and compound Q10-P2 (28.95 mg, purity 97.29%) as a yellow solid.

[0428] Compound Q10-P1:

[0429] LCMS: m / z=612.0 (M+H) + ,Rt=2.541min.

[0430] 1H NMR: (400MHz, CDCl3-d)δ8.04(d,J=8.1Hz,2H),7.66-7.77(m,3H),7.47-7.56(m,1H),7.34-7.42(m,3 H),7.20(dd,J=7.7,1.7Hz,2H),4.96(s,2H),4.62-4.71(m,2H),4.12-4.22(m,3H),3.47-3.54(m,2H).

[0431] Compound Q10-P2:

[0432] LCMS: m / z=612.1 (M+H) + ,Rt=2.542min.

[0433] 1 H NMR: (400MHz, CDCl3-d)δ8.05(d,J=8.1Hz,2H),7.69-7.76(m,3H),7.51(br d,J=3.9Hz,1H),7.34-7.43(m,3H),7.20(br d,J=7.5Hz,2H),4.92(s,2H),4.64-4.72(m,2H),4.15-4.22(m,3H),3.51(t,J=5.8Hz,2H).

[0434] Preparation Example 28: Synthesis of (Z)-4-phenyl-N-(2-sulfamoylethyl)-N'-((4-(trifluoromethyl)phenyl)sulfonyl)-3-(2-(trifluoromethane)thiazol-5-yl)-4,5-dihydro-1H-pyrazole-1-carboximide (S, R enantiomers of Compound Q11)

[0435] To a solution of compound 1 (3.00 g, 15.2 mmol) in DCM (30.0 mL) were added DIEA (5.90 g, 45.6 mmol, 7.95 mL) and HATU (6.94 g, 18.2 mmol). Compound 1a (2.49 g, 25.5 mmol) was then added. The mixture was stirred at 25° C. for 2 hours. TLC (petroleum ether:ethyl acetate=5:1, R f =0.25) indicated that compound 1 was completely consumed and a new spot was formed. The reaction mixture was filtered and concentrated under reduced pressure to obtain a residue. The residue was purified by flash silica gel chromatography ( 40g The product was purified by silica gel flash column with a gradient of 0-20% ethyl acetate / petroleum ether at 45 mL / min as eluent to afford compound 2 (2.20 g, 8.24 mmol, 54.1% yield, 90.0% purity) as a yellow oil.

[0436] 1 H NMR: (400MHz, CDCl3-d) δ8.63 (d, J = 0.9Hz, 1H), 3.81 (s, 3H), 3.40 (s, 3H).

[0437] Compound 2 (2.00 g, 8.33 mmol) was dissolved in THF (20.0 mL) at 0°C and N2. Compound 2a (1 M, 12.4 mL) was then added dropwise at 0°C and N2. The mixture was stirred at 0°C for 2 hours. TLC (petroleum ether:ethyl acetate = 5:1, R f =0.45) indicated that compound 2 was completely consumed and a new spot was formed. The reaction mixture was quenched with saturated aqueous NH4Cl solution (50.0 mL) at 0°C under N2 and then extracted with ethyl acetate (50.0 mL×2). The combined organic layers were washed with brine (50.0 mL), dried over Na2SO4, filtered and concentrated under reduced pressure to obtain a residue. The residue was purified by flash silica gel chromatography ( 20g The product was purified by silica gel flash chromatography (0-20% ethyl acetate / petroleum ether gradient @ 50 mL / min) to afford compound 3 as a yellow oil (1.10 g, 3.65 mmol, 43.8% yield, 90.0% purity).

[0438] LCMS: m / z=272.0 (M+H) + , Rt=1.474min.

[0439] To a solution of compound 3 (1.00 g, 3.69 mmol) and formaldehyde (1.50 g, 18.4 mmol, 1.37 mL) in MeOH (10.0 mL) were added piperidine (313 mg, 3.69 mmol, 364 μL) and AcOH (442 mg, 7.37 mmol, 422 μL). The mixture was stirred at 50° C. for 2 hours. LCMS (Rt=2.829 min) showed that compound 3 was completely consumed, and a major peak with the desired mass was detected. The reaction mixture was filtered and concentrated under reduced pressure to give compound 4 (1.00 g, 3.53 mmol, 95.7% yield) as a yellow oil.

[0440] LCMS: m / z=283.8 (M+H) +, Rt=2.829min.

[0441] N2H4·H2O (176 mg, 3.53 mmol, 171 μL) was dissolved in EtOH (10.0 mL). Compound 4 (1.00 g, 3.53 mmol) dissolved in EtOH (10.0 mL) was then added dropwise to the mixture over 10 minutes. The reaction mixture was stirred at 25°C for 2 hours. LCMS (Rt = 2.787 min) showed that compound 4 was completely consumed, and a main peak with the desired mass number was detected. The reaction mixture was concentrated under reduced pressure to give INT_6 (1.05 g, 3.53 mmol, crude product) as a white solid.

[0442] LCMS: m / z=297.8 (M+H) + , Rt=2.787min.

[0443] Compound 5 (1.00 g, 2.90 mmol) was dissolved in toluene (10.0 mL). INT_6 (861 mg, 2.90 mmol) was then added. The mixture was stirred at 110° C. for 2 hours. LCMS (Rt=2.958 min) showed that INT_6 was completely consumed, and a main peak with the desired mass number was detected. The reaction mixture was filtered and concentrated under reduced pressure to obtain a residue. The residue was purified by preparative HPLC (TFA conditions) (chromatographic column: Phenomenex luna C18 100×40 mm×3 μm; mobile phase: [H2O (0.1% TFA)-ACN]; gradient: 49%-79% B in 10.0 min) to give compound 6 (225 mg, 389 μmol, yield 13.4%, purity 95.0%) as a yellow oil.

[0444] LCMS: m / z=548.5 (M+H) + , Rt=2.958min.

[0445] LCMS: m / z=548.5 (M+H) + , Rt=2.952min.

[0446] Compound 6 (225 mg, 389 μmol) was dissolved in toluene (2.00 mL). POCl (125 mg, 820 μmol, 76.4 μL) and DIEA (106 mg, 820 μmol, 142 μL) were then added. The mixture was stirred at 110° C. for 2 hours. LCMS (Rt=3.127 min) showed that compound 6 was completely consumed, and a main peak with the desired mass number was detected. The reaction mixture was quenched with 1 M NaOH (10.0 mL) at 0° C. and then extracted with ethyl acetate (10.0 mL×3). The combined organic layers were washed with brine (10.0 mL), dried over Na2SO4, filtered and concentrated under reduced pressure to give compound 7 (225 mg, 396 μmol, crude product) as a yellow oil.

[0447] LCMS: m / z=566.5 (M+H) + , Rt=3.127min.

[0448] Compound 7 (225 mg, 396 μmol) was dissolved in DCM (2.00 mL). TEA (200 mg, 1.98 mmol, 276 μL) and compound 7a (95.6 mg, 595 μmol) were then added. The mixture was stirred at 25 ° C for 2 hours. LCMS (Rt = 2.726 min) showed that compound 7 was completely consumed and a major peak with the desired mass number was detected. The reaction mixture was filtered and concentrated under reduced pressure to obtain a residue. The residue was purified by flash silica gel chromatography ( 20g The product was purified by silica gel flash column with a gradient of 0-50% petroleum ether / ethyl acetate at 40 mL / min as the eluent to afford compound 8 (125 mg, 187 μmol, 47.1% yield, 98.09% purity) as a white solid.

[0449] LCMS: m / z=654.4 (M+H) + , Rt=2.726min.

[0450] Compound 8:

[0451] LCMS: m / z=654.4 (M+H) + , Rt=2.721min.

[0452] 1H NMR: (400MHz, CDCl3-d)δ8.05(d,J=8.3Hz,2H),7.71(d,J=8.4Hz,2H),7.64(d,J=0.6Hz,1H),7.46-7.53(m,1H ),7.33-7.42(m,3H),7.18-7.25(m,2H),4.97(s,2H),4.61-4.72(m,2H),4.13-4.22(m,3H),3.49-3.54(m,2H).

[0453] Compound 8 (110 mg) was purified by SFC (chromatographic column: DAICEL CHIRALPAK AD (250 mm × 30 mm, 10 μm); mobile phase: [CO2-EtOH (0.1% NH3H2O)]; B%: 50%, isocratic elution mode) to obtain compound Q11-P1 (44.4 mg, purity 98.09%) as a white solid, and compound Q11-P2 (44.6 mg, purity 100%) as a white solid.

[0454] Compound Q11-P1:

[0455] LCMS: m / z=654.4 (M+H) + , Rt=2.721min.

[0456] 1 H NMR: (400MHz, CDCl3-d)δ8.05(d,J=8.4Hz,2H),7.71(d,J=8.5Hz,2H),7.64(s,1H),7.48(br t,J=5.6Hz,1H),7.34-7.42(m,3H),7.21(dd,J=7.8,1.5Hz,2H),4.93(s,2H),4.61-4.73(m,2H),4.12-4.23(m,3H),3.49-3.53(m,2H).

[0457] Compound Q11-P2:

[0458] LCMS: m / z=654.4 (M+H) + , Rt=2.717 min.

[0459] 1H NMR: (400 MHz, CDCl3-d) δ8.05 (d, J = 8.3 Hz, 2H), 7.71 (d, J = 8.3 Hz, 2H), 7.64 (d, J = 1.0 Hz,1H),7.45-7.52(m,1H),7.33-7.42(m,3H),7.21(dd,J=7.8,1.4 Hz,2H),4.97(s,2H),4.62-4.71(m,2H),4.14-4.21(m,3H),3.48-3.53(m,2H).

[0460] Preparation Example 29: Synthesis of (Z)-3-(2-chlorothiazol-5-yl)-4-phenyl-N-(2-sulfamoylethyl)-N'-((4-(trifluoromethyl)phenyl)sulfonyl)-4,5-dihydro-1H-pyrazole-1-carboximide (S, R enantiomers of Compound Q12)

[0461] To a solution of compound 1a (9.00 g, 92.2 mmol) in DCM (90.0 mL) were added DIEA (24.8 g, 192 mmol, 33.5 mL) and HATU (25.1 g, 66.0 mmol). 2-Chlorothiazole-5-carboxylic acid (9.00 g, 55.0 mmol) was then added. The mixture was stirred at 25°C for 1 hour. TLC (petroleum ether:ethyl acetate = 5:1, R f =0.25) indicated that compound 1 was completely consumed and a new spot was formed. The reaction mixture was filtered and concentrated under reduced pressure to obtain a residue. The residue was purified by flash silica gel chromatography ( 80 g The residue was purified by silica gel flash column (0-15% ethyl acetate / petroleum ether gradient @ 60 mL / min) to afford compound 2 as a yellow oil (7.00 g, 32.1 mmol, 58.4% yield, 95.0% purity).

[0462] 1 H NMR: (400 MHz, CDCl3-d) δ8.30(s,1H),3.77(s,3H),3.36(s,3H).

[0463] Compound 2 (4.00 g, 19.3 mmol) was dissolved in THF (40.0 mL) at 0°C under N2. Compound 2a (1 M, 29.0 mL) was then added dropwise at 0°C under N2. The mixture was stirred at 0°C for 2 hours. TLC (petroleum ether:ethyl acetate = 5:1, R f=0.5) indicated that compound 2 was completely consumed and a new spot was formed. The reaction mixture was quenched with saturated aqueous NH4Cl solution (80.0 mL) at 0°C under N2, and then extracted with ethyl acetate (50.0 mL×2). The combined organic layers were washed with brine (50.0 mL), dried over Na2SO4, filtered, and concentrated under reduced pressure to obtain a residue. The residue was purified by flash silica gel chromatography ( 40 g The product was purified by silica gel flash chromatography (eluent: 0-15% ethyl acetate / petroleum ether gradient @ 50 mL / min) to afford compound 3 as a yellow oil (2.73 g, 10.9 mmol, 56.3% yield, 95.0% purity).

[0464] 1 H NMR: (400 MHz, CDCl3-d) δ8.10(s,1H),7.24-7.39(m,5H),4.14(s,2H).

[0465] To a solution of compound 3 (2.00 g, 8.41 mmol) and formaldehyde (3.41 g, 42.0 mmol, 3.13 mL) in MeOH (20.0 mL) were added piperidine (716 mg, 8.41 mmol, 830 μL) and AcOH (1.01 g, 16.8 mmol, 963 μL). The mixture was stirred at 50° C. for 1 hour. LCMS (Rt=2.645 min) showed that compound 3 was completely consumed, and a major peak with the desired mass was detected. The reaction mixture was filtered and concentrated under reduced pressure to give compound 4 (2.10 g, 8.41 mmol, crude product) as a yellow oil.

[0466] LCMS: m / z=249.8 (M+H) + , Rt=2.645min.

[0467] N2H4·H2O (0.62 g, 12.3 mmol, 600 μL) was dissolved in EtOH (21.0 mL). Compound 4 (2.10 g, 8.41 mmol) dissolved in EtOH (21.0 mL) was then added dropwise to the mixture over 10 minutes. The reaction mixture was stirred at 25°C for 2 hours. LCMS (Rt = 2.425 min) showed that compound 4 was completely consumed, and a main peak with the desired mass number was detected. The reaction mixture was concentrated under reduced pressure to give INT_7 (2.22 g, 8.42 mmol, crude product) as a white solid.

[0468] LCMS: m / z=263.8 (M+H) +, Rt=2.425min.

[0469] Compound 5 (2.80 g, 8.11 mmol) was dissolved in toluene (30.0 mL). INT_7 (2.14 g, 8.11 mmol) was then added. The mixture was stirred at 110° C. for 1 hour. LCMS (Rt=2.844 min) showed that INT_7 was completely consumed, and a main peak with the desired mass number was detected. The reaction mixture was filtered and concentrated under reduced pressure to obtain a residue. The residue was purified by preparative HPLC (TFA conditions) (chromatographic column: Phenomenex luna C18 100×40 mm×3 μm; mobile phase: [H2O (0.1% TFA)-ACN]; gradient: 47%-77% B in 10.0 min) to give compound 6 (300 mg, 524 μmol, yield 6.47%, purity 90.0%) as a yellow oil.

[0470] LCMS: m / z=514.5 (M+H) + , Rt=2.844min.

[0471] LCMS: m / z=514.5 (M+H) + , Rt=2.839min.

[0472] Compound 6 (150 mg, 291 μmol) was dissolved in toluene (1.50 mL). POCl (89.3 mg, 582 μmol, 54.3 μL) and DIEA (75.3 mg, 582 μmol, 101 μL) were then added. The mixture was stirred at 110° C. for 1 hour. LCMS (Rt=3.033 min) showed that compound 6 was completely consumed, and a main peak with the desired mass number was detected. The reaction mixture was quenched with 1 M NaOH (10.0 mL) at 0° C. and then extracted with ethyl acetate (10.0 mL×3). The combined organic layers were washed with brine (10.0 mL), dried over Na2SO4, filtered and concentrated under reduced pressure to give compound 7 (150 mg, 281 μmol, crude product) as a yellow oil.

[0473] LCMS: m / z=532.5 (M+H) + , Rt=3.033min.

[0474] Compound 7 (150 mg, 281 μmol) was dissolved in DCM (1.50 mL). TEA (142 mg, 1.41 mmol, 195 μL) and compound 4a (67.7 mg, 421 μmol) were then added. The mixture was stirred at 25 ° C for 2 hours. LCMS (Rt = 2.598 min) showed that compound 7 was completely consumed and a major peak with the desired mass number was detected. The reaction mixture was filtered and concentrated under reduced pressure to obtain a residue. The residue was purified by flash silica gel chromatography ( 12g The product was purified by silica gel flash column with a gradient of 0-50% petroleum ether / ethyl acetate at 40 mL / min as the eluent to afford compound 8 (48.0 mg, 75.8 μmol, 26.9% yield, 98.16% purity) as a yellow solid.

[0475] LCMS: m / z=620.4 (M+H) + , Rt=2.598min.

[0476] Compound 8:

[0477] LCMS: m / z=620.5 (M+H) + , Rt=2.598min.

[0478] 1 H NMR: EC25148-257-P1N2 (400MHz, CDCl3-d) δ8.04 (br d, J=8.1Hz, 2H), 7.66-7.74 (m, 2H), 7.45 (br s,1H),7.32-7.39(m,3H),7.28(d,J=3.3Hz,1H),7.15-7.21(m,2H),5.00(br s,2H),4.56-4.67(m,2H),4.07-4.18(m,3H),3.47-3.53(m,2H).

[0479] Compound 8 (40.0 mg) was purified by SFC (chromatographic column: DAICELCHIRALPAKAD (250 mm×30 mm, 10 μm); mobile phase: [CO2-EtOH:ACN=7:3 (0.1% NH3H2O)]; B%: 45%, isocratic elution mode) to obtain compound Q12-P1 (15.10 mg, purity 98.54%) as a yellow solid, and compound Q12-P2 (14.76 mg, purity 98.67%) as a yellow solid.

[0480] Compound Q12-P1:

[0481] LCMS: m / z=620.4 (M+H) + , Rt=2.595min.

[0482] 1 H NMR: (400MHz, CDCl3-d)δ8.04(d,J=8.1Hz,2H),7.70(d,J=8.3Hz,2H),7.46(br s,1H),7.33-7.40(m,3H),7.28(s,1H),7.19(br d,J=6.8Hz,2H),5.03(s,2H),4.56-4.67(m,2H),4.08-4.18(m,3H),3.50(t,J=5.9Hz,2H).

[0483] Compound Q12-P2:

[0484] LCMS: m / z=620.4 (M+H) + , Rt=2.595min.

[0485] 1 H NMR: (400MHz, CDCl3-d) δ8.04(br d,J=8.3Hz,2H),7.70(br d,J=8.1Hz,2H),7.45(br s,1H),7.33-7.40(m,3H),7.28(s,1H),7.19(br d,J=6.9Hz,2H),5.02(s,2H),4.55-4.67(m,2H),4.08-4.18(m,3H),3.50(br t,J=5.8Hz,2H).

[0486] Preparation Example 30: Synthesis of (Z)-3-(5-chloropyridin-2-yl)-4-phenyl-N-(2-sulfamoylethyl)-N'-((4-(trifluoromethyl)phenyl)sulfonyl)-4,5-dihydro-1H-pyrazole-1-carboximide (S, R enantiomers of compound Q13)

[0487] To a solution of compound 1 (5.00 g, 31.7 mmol) in DCM (50.0 mL) were added DIEA (10.2 g, 79.3 mmol, 13.8 mL) and HATU (14.4 g, 38.0 mmol). Compound 1a (5.20 g, 53.3 mmol) was then added. The mixture was stirred at 25° C. for 1 hour. TLC (petroleum ether:ethyl acetate=5:1, R f =0.3) indicated that compound 1 was completely consumed and a new spot was formed. The reaction mixture was filtered and concentrated under reduced pressure to obtain a residue. The residue was purified by flash silica gel chromatography ( 40g The product was purified by silica gel flash column with a gradient of 0-15% ethyl acetate / petroleum ether at 40 mL / min as eluent to afford compound 2 (4.35 g, 21.6 mmol, 68.3% yield, 90.0% purity) as a yellow oil.

[0488] 1 H NMR: (400MHz, CDCl3-d) δ 8.57 (d, J = 2.1 Hz, 1H), 7.76 (dd, J = 8.3, 2.3 Hz, 1H), 7.65 (br s, 1H), 3.75 (br s, 3H), 3.40 (br s, 3H).

[0489] Compound 2 (1.00 g, 4.98 mmol) was dissolved in THF (10.0 mL) at 0°C under N2. Compound 2a (1 M, 7.48 mL) was then added dropwise at 0°C under N2. The mixture was stirred at 0°C for 2 hours. TLC (petroleum ether:ethyl acetate = 5:1, R f =0.45) indicated that compound 2 was completely consumed and a new spot was formed. The reaction mixture was quenched with saturated aqueous NH4Cl solution (20.0 mL) at 0°C under N2 and then extracted with ethyl acetate (20.0 mL×2). The combined organic layers were washed with brine (20.0 mL), dried over Na2SO4, filtered and concentrated under reduced pressure to obtain a residue. The residue was purified by flash silica gel chromatography ( 10g The product was purified by silica gel flash chromatography (0-10% ethyl acetate / petroleum ether gradient @ 30 mL / min) to afford compound 3 as a yellow oil (382 mg, 1.15 mmol, yield 23.1%, purity 70.0%).

[0490] 1 H NMR: (400MHz, CDCl3-d) δ8.59 (d, J=2.3Hz, 1H), 7.94 (d, J=8.4Hz, 1H), 7.72 (dd, J=8.4, 1.9Hz, 1H), 7.21-7.30 (m, 5H), 4.43 (s, 2H).

[0491] To a solution of compound 3 (382 mg, 1.65 mmol) and formaldehyde (247 mg, 8.24 mmol, 227 μL) in MeOH (4.00 mL) were added piperidine (140 mg, 1.65 mmol, 162 μL) and AcOH (198 mg, 3.30 mmol, 188 μL). The mixture was stirred at 50° C. for 1 hour. LCMS (Rt=2.829 min) showed that compound 3 was completely consumed, and a major peak with the desired mass was detected. The reaction mixture was filtered and concentrated under reduced pressure to give compound 4 (400 mg, 1.64 mmol, crude product) as a yellow oil.

[0492] LCMS: m / z=244.0 (M+H) + , Rt=1.394min.

[0493] Compound 4 (200 mg, 820 μmol) and N₂H₄·H₂O (97.0 mg, 1.94 mmol, 93.9 μL) were dissolved in EtOH (2.0 mL). The reaction mixture was stirred at 25°C for 2 hours. LCMS (Rt = 1.218 min) showed complete consumption of compound 4, with a major peak of the desired mass detected. The reaction mixture was concentrated under reduced pressure to afford INT_8 (200 mg, 776 μmol, crude) as a white solid.

[0494] LCMS: m / z=258.0 (M+H) + , Rt=1.218min.

[0495] Compound 5 (250 mg, 724 μmol) was dissolved in toluene (2.50 mL). INT_8 (186 mg, 724 μmol) was then added. The mixture was stirred at 110 ° C for 1 hour. LCMS (Rt = 1.673 min) showed that INT_8 was completely consumed and a main peak with the required mass number was detected. The reaction mixture was filtered and concentrated under reduced pressure to obtain a residue. The residue was purified by preparative HPLC (TFA conditions) (chromatographic column: CD12-WePure Biotech Phenyl-Hexyl 150×25mm×7μm; mobile phase: [H2O (0.1% TFA)-ACN]; gradient: 48%-78% B in 10.0 min) to give a yellow solid compound 6 (98.0 mg, 192 μmol, yield 25.2%, purity 95.2%).

[0496] LCMS: m / z=509.0 (M+H) + , Rt=1.673min.

[0497] LCMS: m / z=509.0 (M+H) + , Rt=1.671min.

[0498] Compound 6 (98.0 mg, 192 μmol) was dissolved in toluene (1.00 mL). POCl (59.0 mg, 385 μmol, 35.9 μL) and DIEA (49.7 mg, 385 μmol, 67.0 μL) were then added. The mixture was stirred at 110° C. for 1 hour. LCMS (Rt=1.800 min) showed that compound 6 was completely consumed, and a main peak with the desired mass number was detected. The reaction mixture was quenched with 1 M NaOH (10.0 mL) at 0° C. and then extracted with ethyl acetate (10.0 mL×3). The combined organic layers were washed with brine (10.0 mL), dried over Na2SO4, filtered and concentrated under reduced pressure to give compound 7 (101 mg, 191 μmol, crude product) as a yellow oil.

[0499] LCMS: m / z=527.0 (M+H) + , Rt=1.800min.

[0500] Compound 7 (101 mg, 191 μmol) was dissolved in DCM (1.00 mL). TEA (96.9 mg, 957 μmol, 133 μL) and compound 7a (46.1 mg, 287 μmol) were then added. The mixture was stirred at 25° C. for 2 hours. LCMS (Rt=1.579 min) showed that compound 7 was completely consumed, and a major peak with the desired mass number was detected. The reaction mixture was filtered and concentrated under reduced pressure to obtain a residue. The residue was purified by flash silica gel chromatography ( 12g The product was purified by silica gel flash column with a gradient of 0-50% petroleum ether / ethyl acetate at 40 mL / min as the eluent to afford compound 8 (44.5 mg, 72.0 μmol, 37.7% yield, 97.59% purity) as a white solid.

[0501] LCMS: m / z=615.0 (M+H) + , Rt=1.579min.

[0502] Compound 8:

[0503] LCMS: m / z=614.5 (M+H) + , Rt=2.735min.

[0504] 1H NMR: (400MHz, CDCl3-d)δ8.42(d,J=2.1Hz,1H),8.05(d,J=8.1Hz,2H),7.83(d,J=8.5Hz,1H),7.61-7.72(m,4H),7.19-7.26(m,2H), 7.15(d,J=6.9Hz,2H),5.16(s,2H),4.92(dd,J=11.4,4.8Hz,1H),4.47(t,J=11.6Hz,1H),4.15-4.26(m,3H),3.55(t,J=6.0Hz,2H).

[0505] Compound 8 (40.0 mg) was purified by SFC (chromatographic column: DAICELCHIRALPAK AS (250 mm×30 mm, 10 μm); mobile phase: [CO2-MeOH (0.1% NH3H2O)]; B%: 45%, isocratic elution mode) to obtain compound Q13-P1 (15.00 mg, purity 99.06%) as a white solid, and compound Q13-P2 (15.40 mg, purity 100%) as a white solid.

[0506] Compound Q13-P1:

[0507] LCMS: m / z=614.5 (M+H) + , Rt=2.700min.

[0508] 1 H NMR: (400MHz, CDCl3-d) δ8.43(d,J=1.9Hz,1H),8.05(d,J=8.3Hz,2H),7.84(d,J=8.4Hz,1H),7.70(br d,J=8.3Hz,2H),7.64(br dd,J=8.4,2.4Hz,2H),7.23(br d,J=7.0Hz,2H),7.15(br d,J=7.0Hz,2H),5.06(s,2H),4.93(dd,J=11.4,4.7Hz,1H),4.47(t,J=11.6Hz,1H),4.14-4.27(m,3H),3.55(t,J=6.0Hz,2H).

[0509] Compound Q13-P2:

[0510] LCMS: m / z=614.5 (M+H) + , Rt=2.700min.

[0511] 1H NMR: (400MHz, CDCl3-d) δ8.43(d,J=1.8Hz,1H),8.05(d,J=8.1Hz,2H),7.84(d,J=8.5Hz,1H),7.70(d,J=8.3Hz,2H),7.64(br dd,J=8.5,2.4Hz,2H),7.23(br d,J=7.0Hz,2H),7.12-7.20(m,2H),5.07(s,2H),4.93(dd,J=11.4,4.8Hz,1H),4.47(t,J=11.6Hz,1H),4.15-4.27(m,3H),3.55(t,J=6.0Hz,2H).

[0512] Preparation Example 31: Synthesis of (Z)-4-phenyl-N-(2-sulfamoylethyl)-N'-((4-(trifluoromethyl)phenyl)sulfonyl)-3-(5-(trifluoromethane)pyridin-2-yl)-4,5-dihydro-1H-pyrazole-1-carboximide (S, R enantiomers of compound Q14)

[0513] To a solution of compound 1 (5.00 g, 26.1 mmol) in DCM (50.0 mL) were added DIEA (8.45 g, 65.4 mmol, 11.3 mL) and HATU (11.9 g, 31.4 mmol). Compound 1a (4.29 g, 43.9 mmol) was then added. The mixture was stirred at 25° C. for 1 hour. TLC (petroleum ether:ethyl acetate=5:1, R f =0.25) indicated that compound 1 was completely consumed and a new spot was formed. The reaction mixture was filtered and concentrated under reduced pressure to obtain a residue. The residue was purified by flash silica gel chromatography ( 40g The product was purified by silica gel flash column with a gradient of 0-15% ethyl acetate / petroleum ether at 45 mL / min as eluent to afford compound 2 (3.10 g, 13.2 mmol, 50.6% yield, 90.0% purity) as a yellow oil.

[0514] 1 H NMR: (400MHz, CDCl3-d) δ 8.88 (s, 1H), 8.03 (br d, J = 8.0Hz, 1H), 7.65-7.89 (m, 1H), 3.74 (br s, 3H), 3.40 (br s, 3H).

[0515] Compound 2 (1.50 g, 6.41 mmol) was dissolved in THF (15.0 mL) at 0°C and N2. Compound 2a (1 M, 9.61 mL) was then added dropwise at 0°C and N2. The mixture was stirred at 0°C for 2 hours. TLC (petroleum ether:ethyl acetate = 5:1, R f =0.45) indicated that compound 2 was completely consumed and a new spot was formed. The reaction mixture was quenched with saturated aqueous NH4Cl solution (30.0 mL) at 0°C under N2 and then extracted with ethyl acetate (30.0 mL×2). The combined organic layers were washed with brine (30.0 mL), dried over Na2SO4, filtered and concentrated under reduced pressure to obtain a residue. The residue was purified by flash silica gel chromatography ( 20g Purification by silica gel flash chromatography (eluting with 0-10% ethyl acetate / petroleum ether gradient @ 40 mL / min) afforded compound 3 as a yellow oil (1.05 g, 3.7 mmol, yield 58.7%, purity 95.0%).

[0516] 1 H NMR: (400MHz, CDCl3-d) δ8.99 (d, J = 0.6Hz, 1H), 8.03-8.21 (m, 2H), 7.23-7.36 (m, 5H), 4.56 (s, 2H).

[0517] To a solution of compound 3 (1.00 g, 3.77 mmol) and formaldehyde (1.53 g, 18.8 mmol, 1.40 mL) in MeOH (10.0 mL) were added piperidine (321 mg, 3.77 mmol, 372 μL) and AcOH (452 ​​mg, 7.54 mmol, 431 μL). The mixture was stirred at 50° C. for 1 hour. LCMS (Rt=2.684 min) showed that compound 3 was completely consumed, and a major peak with the desired mass was detected. The reaction mixture was filtered and concentrated under reduced pressure to give compound 4 (627 mg, 2.26 mmol, crude product) as a yellow oil.

[0518] LCMS: m / z=277.9 (M+H) + , Rt=2.684min.

[0519] N2H4·H2O (110 mg, 2.18 mmol, 106 μL) was dissolved in EtOH (6.00 mL). Compound 4 (600 mg, 2.16 mmol) dissolved in EtOH (6.0 mL) was then added dropwise to the mixture over 10 minutes. The reaction mixture was stirred at 25°C for 2 hours. LCMS (Rt = 2.495 min) showed that compound 4 was completely consumed, and a main peak with the desired mass number was detected. The reaction mixture was concentrated under reduced pressure to give INT_9 (630 mg, 2.16 mmol, crude product) as a white solid.

[0520] LCMS: m / z=291.9 (M+H) + , Rt=2.495min.

[0521] Compound 5 (700 mg, 2.03 mmol) was dissolved in toluene (7.00 mL). INT_9 (590 mg, 2.03 mmol) was then added. The mixture was stirred at 110° C. for 1 hour. LCMS (Rt=2.945 min) showed that INT_9 was completely consumed and a main peak with the desired mass number was detected. The reaction mixture was filtered and concentrated under reduced pressure to obtain a residue. The residue was purified by preparative HPLC (TFA conditions) (chromatographic column: Phenomenex luna C18 100×40 mm×3 μm; mobile phase: [H2O (0.1% TFA)-ACN]; gradient: 50%-80% B in 10.0 min) to give compound 6 (295 mg, 489 μmol, yield 24.1%, purity 90.0%) as a yellow oil.

[0522] LCMS: m / z=542.6 (M+H) + , Rt=2.945min.

[0523] LCMS: m / z=542.6 (M+H) + , Rt=2.943min.

[0524] Compound 6 (180 mg, 331 μmol) was dissolved in toluene (2.00 mL). POCl (101 mg, 663 μmol, 61.8 μL) and DIEA (85.7 mg, 663 μmol, 115 μL) were then added. The mixture was stirred at 110° C. for 1 hour. LCMS (Rt=3.104 min) showed that compound 6 was completely consumed, and a main peak with the desired mass number was detected. The reaction mixture was quenched with 1 M NaOH (10.0 mL) at 0° C. and then extracted with ethyl acetate (10.0 mL×3). The combined organic layers were washed with brine (10.0 mL), dried over Na2SO4, filtered and concentrated under reduced pressure to give compound 7 (180 mg, 320 μmol, crude product) as a yellow oil.

[0525] LCMS: m / z=560.5 (M+H) + , Rt=3.104min

[0526] Compound 7 (180 mg, 320 μmol) was dissolved in DCM (2.00 mL). TEA (162 mg, 1.60 mmol, 223 μL) and compound 7a (77.3 mg, 481 μmol) were then added. The mixture was stirred at 25 ° C for 2 hours. LCMS (Rt = 2.750 min) showed that compound 7 was completely consumed and a major peak with the desired mass number was detected. The reaction mixture was filtered and concentrated under reduced pressure to obtain a residue. The residue was purified by flash silica gel chromatography ( 12g The product was purified by silica gel flash column with a gradient of 0-50% petroleum ether / ethyl acetate at 30 mL / min as the eluent to afford compound 8 (125 mg, 191 μmol, 59.6% yield, 99.36% purity) as a white solid.

[0527] LCMS: m / z=648.5 (M+H) + , Rt=2.750min.

[0528] Compound 8:

[0529] LCMS: m / z=648.5 (M+H) + , Rt=2.748min.

[0530] 1H NMR: (400MHz, CDCl3-d)δ8.73(s,1H),8.01-8.09(m,3H),7.89(dd,J=8.4,1.9Hz,1H),7.70(d,J=8.1Hz,3H),7.27-7.31(m,1H),7.20-7.2 5(m,1H),7.15-7.20(m,2H),5.11(s,2H),4.97(dd,J=11.5,4.9Hz,1H),4.50(t,J=11.7Hz,1H),4.16-4.30(m,3H),3.56(t,J=6.1Hz,2H).

[0531] Compound 8 (90.0 mg) was purified by SFC (chromatographic column: DAICELCHIRALPAKAD (250 mm×30 mm, 10 μm); mobile phase: [CO2-IPA (0.1% NH3H2O)]; B%: 30%, isocratic elution mode) to obtain compound Q14-P1 (23.73 mg, purity 99.85%) as a white solid, and compound Q14-P2 (23.93 mg, purity 100%) as a white solid.

[0532] Compound Q14-P1:

[0533] LCMS: m / z=648.5 (M+H) + , Rt=2.745min.

[0534] 1 H NMR: (400MHz, CDCl3-d)δ8.73(s,1H),8.01-8.10(m,3H),7.89(dd,J=8.4,1.9Hz,1H),7.71(d,J=8.4Hz,3H),7.28-7.30(m,1H),7.23(d,J= 7.0Hz,1H),7.15-7.20(m,2H),5.05(s,2H),4.98(dd,J=11.4,4.8Hz,1H),4.50(t,J=11.6Hz,1H),4.18-4.29(m,3H),3.56(t,J=6.0Hz,2H).

[0535] Compound Q14-P2:

[0536] LCMS: m / z=648.5 (M+H) + , Rt=2.745min.

[0537] 1H NMR: (400MHz, CDCl3-d) δ8.73 (s, 1H), 8.01-8.09 (m, 3H), 7.90 (dd, J = 8.4, 1.9Hz, 1H), 7.71 (br d,J=8.4Hz,3H),7.28-7.31(m,1H),7.20-7.24(m,1H),7.17(d,J=6.9Hz,2H),5.04(s,2H),4. 98 (dd, J = 11.6, 4.8 Hz, 1H), 4.50 (t, J = 11.7 Hz, 1H), 4.19-4.29 (m, 3H), 3.56 (t, J = 5.9 Hz, 2H).

[0538] Preparation Example 32: Synthesis of S and R enantiomers of compound Q15

[0539] A solution of compound 1 (300 mg, 708 μmol, 1.00 eq.) in TFA (2.00 mL) was stirred at 90°C for 1 hour. LC-MS showed complete consumption of compound 1. The reaction mixture was filtered and concentrated under reduced pressure to afford compound INT_2 (70.0 mg, crude) as a brown oil for later use.

[0540] To a solution of INT_1 (250 mg, 1.08 mmol, 1.00 eq.) and TEA (274 mg, 2.70 mmol, 376 μL, 2.50 eq.) in DCM (2.00 mL) was added compound 1a (203 mg, 1.30 mmol, 163 μL, 1.20 eq.) in portions at 0°C. The mixture was stirred at 0°C for 1 hour. LCMS (product Rt = 2.455 min) showed that INT_1 was completely consumed, and a major peak with the desired mass was detected. The reaction mixture was filtered and concentrated under reduced pressure to give compound 2 (379 mg, crude product) as a brown oil.

[0541] LCMS: m / z=352.0 (M+H) + ,Rt=2.455min.

[0542] To a solution of compound 2 (379 mg, 1.08 mmol, 1.00 eq) in toluene (2.00 mL) was added compound 2a (277 mg, 1.08 mmol, 1.00 eq). The mixture was stirred at 110° C. for 1 hour. LCMS (product Rt=2.957 min) showed that compound 2 was completely consumed, and a major peak with the desired mass was detected. The reaction mixture was filtered and concentrated under reduced pressure to give compound 3 (554 mg, crude product) as a brown oil.

[0543] LCMS: m / z=514.0 (M+H) + ,Rt=2.957min.

[0544] To a solution of compound 3 (554 mg, 1.08 mmol, 1.00 eq.) in toluene (2.00 mL) were added POCl (330 mg, 2.16 mmol, 201 μL, 2.00 eq.) and DIEA (279 mg, 2.16 mmol, 376 μL, 2.0 eq.). The mixture was stirred at 110° C. for 1 hour. LCMS (product Rt=3.136 min) showed that compound 3 had been completely consumed, and a major peak with the desired mass was detected. The reaction mixture was quenched by the addition of 1.00 M NaOH (20.0 mL) at 20° C., followed by extraction with ethyl acetate (30.0 mL×3). The combined organic layers were washed with brine (30.0 mL), dried over NaSO, filtered, and concentrated under reduced pressure to give compound 4 (574 mg, crude product) as a brown oil.

[0545] LCMS: m / z=532.0 (M+H) + ,Rt=3.136min.

[0546] To a solution of compound 4 (573 mg, 1.08 mmol, 1.00 eq.) in DCM (2.00 mL) was added TEA (544 mg, 5.38 mmol, 749 μL, 5.00 eq.) and INT-2 (68.7 mg, 558 μmol). The mixture was stirred at 25°C for 1 hour. LCMS (product Rt = 2.405 min) showed complete consumption of compound 4, and a major peak with the desired mass was detected. The reaction mixture was filtered and concentrated under reduced pressure to obtain a residue. The residue was purified by preparative HPLC (column: CD02-Waters Xbridge BEH C18 100 × 25 mm × 10 μm; mobile phase: [H2O (0.05% NH3H2O)-ACN]; gradient: 45% to 65% B over 10.0 minutes) to obtain Q15 (65.0 mg, 9.40% yield, 96.3% purity) as a white solid.

[0547] LCMS: m / z=619.2 (M+H) + ,Rt=1.351mins.

[0548] HPLC: product Rt=4.508 mins.

[0549] 1H NMR: (400MHz, CDCl3-d)δ8.24-8.33(m,1H),7.52-7.58(m,2H),7.48-7.52( m,1H),7.28-7.37(m,6H),7.15(d,J=6.8Hz,2H),4.76-4.78(m,3H),4.72(br s,1H),4.56-4.64(m,1H),4.13-4.25(m,3H),3.49-3.60(m,2H).

[0550] 19 F NMR: (376MHz, CDCl3-d) δ-55.723 (s, 3F).

[0551] Q15 was purified by SFC (column: SFC-OX-30-DAICELCHIRALCELOX (250 mm×30 mm, 10 μm); mobile phase: [CO2-EtOH:CAN=7:3 (0.1% NH3H2O)]; B%: 40%, isocratic elution mode) to give white solid Q15-P1 (25.0 mg, 36.6% yield, 95.2% purity) and white solid Q15-P2 (28.0 mg, 40.9% yield, 95.0% purity).

[0552] Compound Q15-P1:

[0553] LCMS: m / z=619.0 (M+H) + ,Rt=1.798min.

[0554] HPLC: product Rt=4.235 min.

[0555] SFC: Product Rt = 0.750 min.

[0556] 1 H NMR: (400MHz, CDCl3-d)δ8.30(br t,J=5.0Hz,1H),7.46-7.63(m,3H),7.28-7.36(m,4H),7.25(s,2H),7.14(d,J=6.8Hz,2H), 4.74(dd,J=11.4,5.3Hz,1H), 4.59(t,J=11.4Hz,1H), 4.12-4.24(m,3H), 3.46-3.64(m,2H).

[0557] 19 F NMR: (376MHz, CDCl3-d) δ-55.716 (s, 3F).

[0558] Compound Q15-P2:

[0559] LCMS: m / z=619.0 (M+H) + ,Rt=1.794min.

[0560] HPLC: product Rt=2.966 min.

[0561] SFC: Product Rt = 1.216 min.

[0562] 1 H NMR: (400MHz, CDCl3-d) δ8.22-8.38(m,1H),7.53-7.60(m,2H),7.50(br d,J=3.5Hz,1H),7.28-7.39(m,4H),7.25(br s,2H),7.14(br d,J=7.1Hz,2H),4.74(dd,J=11.4,5.3Hz,1H),4.50-4.66(m,1H),4.07-4.28(m,3H),3.46-3.68(m,2H).

[0563] 19 F NMR: (376MHz, CDCl3-d) δ-55.716 (s, 3F).

[0564] Preparation Example 33: Synthesis of S and R enantiomers of compound Q16

[0565] To a solution of compound 7 (310 mg, 587 μmol) of Preparation Example 30 in DCM (3.00 mL) was added TEA (297 mg, 2.94 mmol, 409 μL) and INT_2 (72.4 mg, 587 μmol) of Preparation Example 32. The mixture was stirred at 25 ° C for 2 hours. LCMS (Rt = 2.294 minutes) showed that compound 7 was completely consumed, and a main peak with the desired mass was detected. Thin layer chromatography (petroleum ether: ethyl acetate = 1: 1, R f =0.1). The reaction mixture was filtered and concentrated under reduced pressure to obtain a residue. The residue was purified by flash silica gel chromatography ( 20g The product was purified by silica gel flash column with 0-60% petroleum ether:ethyl acetate gradient @ 40 mL / min as eluent to afford compound 1 (44.6 mg, 72.6 μmol, 12.3% yield, 92.46% purity) as a white solid.

[0566] LCMS: m / z=614.1 (M+H) + ,Rt=2.294min.

[0567] Compound 1:

[0568] LCMS: m / z=614.1 (M+H) + ,Rt=2.302min.

[0569] 1 H NMR: (400MHz, CDCl3-d)δ8.41(d,J=2.4Hz,1H),8.22(br t,J=5.4Hz,1H),8.05(d,J=8.3Hz,2H),7.87(d,J=8.5Hz,1H),7.69(d,J=8.3Hz,2H),7.62(dd,J=8.6,2.4Hz,1H),7.28(br d,J=2.4Hz,1H),7.19-7.26(m,2H),7.12-7.19(m,2H),4.92(dd,J=11.4,4. 7Hz, 1H), 4.44 (t, J = 11.6Hz, 1H), 4.14-4.28 (m, 3H), 3.57 (t, J = 5.8Hz, 2H).

[0570] Q16 (42.5 mg) was purified by SFC (column: SFC-AS-30-DAICELCHIRALPAKAS (250 mm×30 mm, 10 μm); mobile phase: [CO2-MeOH (0.1% NH3H2O)]; B%: 35%, isocratic elution mode) to give a white solid Q16-P1 (15.7 mg, 92.46% purity) and a white solid Q16-P2 (15.9 mg, 95.16% purity).

[0571] Q16-P1:

[0572] LCMS: m / z=614.1 (M+H) + ,Rt=2.288min.

[0573] 1H NMR: (400MHz, CDCl3-d)δ8.41(d,J=2.4Hz,1H),8.18-8.30(m,1H),8.05(d,J=8.3Hz,2 H),7.87(d,J=8.5Hz,1H),7.69(d,J=8.3Hz,2H),7.63(dd,J=8.5,2.3Hz,1H),7.28(br s,1H),7.20-7.25(m,2H),7.15(br d, J=7.3Hz, 2H), 4.93 (dd, J=11.5, 4.6Hz, 1H), 4.44 (t, J=11.6Hz, 1H), 4.15-4.26 (m, 3H), 3.55 (t, J=5.7Hz, 2H).

[0574] Q16-P2:

[0575] LCMS::m / z=648.5(M+H) + ,Rt=2.745min.

[0576] 1H NMR: (400MHz, CDCl3-d) δ8.41(s,1H),8.23(br t,J=5.4Hz,1H),8.05(d,J=8.1Hz,2H),7.86(d,J=8.5Hz,1H),7.68(br d,J=8.1Hz,2H),7.58-7.65(m,1H),7.28(br s,1H),7.20-7.26(m,2H),7.12-7.20(m,2H),4.92(dd,J=11.5,4.5Hz,1H),4.44(t,J=11.5Hz,1H),4.14-4.26(m,3H),3.56(br t,J=5.6Hz,2H).

[0577] Preparation Example 34: Synthesis of S and R enantiomers of compound Q17

[0578] To a solution of compound 8 (300 mg, 563 μmol) of Preparation Example 19 in DCM (3.00 mL) was added TEA (285 mg, 2.82 mmol, 392 μL) and INT_2 (69.4 mg, 563 μmol) of Preparation Example 32. The mixture was stirred at 25 ° C for 2 hours. LCMS (Rt = 2.346 minutes) showed that compound 8 was completely consumed, and a main peak with the desired mass was detected. Thin layer chromatography (petroleum ether: ethyl acetate = 1: 1, R f =0.1). The reaction mixture was filtered and concentrated under reduced pressure to obtain a residue. The residue was purified by flash silica gel chromatography ( 20g The product was purified by silica gel flash column with a gradient of 0-70% petroleum ether:ethyl acetate at 40 mL / min as the eluent to afford compound 1 as a white solid (95.5 mg, 154 μmol, 25.7% yield, 94.09% purity).

[0579] LCMS: m / z=619.0 (M+H) + ,Rt=2.346min.

[0580] Compound 1:

[0581] LCMS: m / z=619.1 (M+H) + ,Rt=2.326min.

[0582] 1 H NMR: (400MHz, CDCl3-d)δ8.15(br t,J=5.5Hz,1H),8.04(d,J=8.3Hz,2H),7.68(d,J=8.3Hz,2H),7.28-7.37(m,3H),7.17(d,J=6.8Hz,2H),6. 64-6.73(m,2H),4.51-4.64(m,2H),4.15(q,J=5.7Hz,2H),4.05(dd,J=10.8,5.7Hz,1H),3.49-3.54(m,2H).

[0583] Q17 (90.0 mg) was purified by SFC (column: SFC-OD-30-DAICEL CHIRALCELOD (250 mm×30 mm, 10 μm); mobile phase: [CO2-MeOH (0.1% NH3H2O)]; B%: 45%, isocratic elution mode) to give a white solid Q17-P1 (16.3 mg, 95.73% purity) and a white solid Q17-P2 (16.6 mg, 96.81% purity).

[0584] Q17-P1:

[0585] LCMS: m / z=619.1 (M+H) + ,Rt=2.344min.

[0586] 1H NMR: (400MHz, CDCl3-d)δ8.15(br t,J=5.7Hz,1H),8.00-8.06(m,2H),7.68(d,J=8.3Hz,2H),7.28-7.37(m,3H),7.13-7.20(m,2H),6.65 -6.71(m,2H),4.51-4.63(m,2H),4.12-4.20(m,2H),4.05(dd,J=10.8,5.6Hz,1H),3.48-3.58(m,2H).

[0587] Q17-P2:

[0588] LCMS: m / z=619.0 (M+H) + ,Rt=2.345min.

[0589] 1 H NMR: (400MHz, CDCl3-d) δ8.15(br t,J=5.6Hz,1H),8.00-8.06(m,2H),7.68(br d,J=8.4Hz,2H),7.29-7.37(m,3H),7.17(br d,J=6.4Hz,2H),6.64-6.72(m,2H),4.51-4.64(m,2H),4.12-4.21(m,2H),4.05(dd,J=10.8,5.6Hz,1H),3.47-3.59(m,2H).

[0590] Example 1. In vitro CB1 receptor inhibitory activity detection experiment based on HTRF-cAMP assay

[0591] The inhibitory effect of the compounds of the present invention on CB1 receptors was analyzed by measuring the changes in intracellular cAMP levels using HTRF (homogeneous time-resolved fluorescence)-cAMP detection method.

[0592] (1) Test compound and control compound

[0593] In this example, the compounds of the present invention Q1-P2, Q2-P2, Q3-P2, Q4-P1, Q5-P1, Q6-P1, Q7-P1, Q8-P2, Q9-P1, Q10-P1, Q11-P1, Q12-P1, Q13-P1, Q14-P2, Q15-P2, Q16-P1 and Q17-P2 were tested; the solvent dimethyl sulfoxide (DMSO; purchased from Sigma) was used as a negative control, and the following control compound 1 and control compound 2 were used as positive controls: control compound 1 was rimonabant (CAS No.: 168273-06-1, purchased from MedChemExpress); control compound 2 was compound 9 disclosed in WO2023196556A1 (synthesized according to the method described on page 73 of the WO2023196556A1 specification).

[0594] (2) Cell construction, culture and passage

[0595] Construction of a CHO cell line stably expressing hCB1 receptor: A CHO stable cell line overexpressing human CB1 was constructed using the pCDNA5 / FRT / TO system. Flp-In-CHO cells (CHO cell line purchased from ATCC) were plated at 6×10 5 Cells were seeded into 6-well plates and incubated with complete medium (F12 + 10% FBS) at 37°C under 5% CO2 overnight. TM Following the instructions for the 3000 Transfection Kit (purchased from Thermo Fisher), a total of 4 μg of a plasmid containing the human CB1 gene (gene information: NM_016083) was transfected into Flp-In-CHO cells. After 6 hours, the medium was changed. The transfected cells were screened for resistance and then treated with hygromycin for 2-3 weeks to obtain a cell pool stably expressing human CB1. Gene expression was confirmed in the cell pool by PCR. Functional analysis (HTRF cAMP assay) was performed on the human CB1-CHO cells after expression was confirmed, resulting in a stable cell line suitable for compound screening.

[0596] For cell culture, a CHO cell line stably expressing the hCB1 receptor was cultured in F-12 medium (117650054, purchased from Gibco) containing 10% fetal bovine serum (76294-180, purchased from Avantor) and 0.2 mg / mL hygromycin B (H8080-1g, purchased from Solarbio) at a temperature of 37°C and a carbon dioxide concentration of 5%.

[0597] For cell passaging, remove old culture medium and wash once with PBS, then add 1 mL of TripLE TMExpress solution and incubate at 37°C for 2 minutes. When the cells detach from the bottom of the dish, add approximately 5 mL of complete culture medium preheated at 37°C. Gently pipette the cell suspension to separate the aggregated cells, transfer the cell suspension to a sterile centrifuge tube, and centrifuge at 1000 rpm for 5 minutes. To maintain cell physiological activity, the cell confluency of the experimental cells was controlled at approximately 80%. Unless otherwise specified, cell recovery and freezing were performed according to conventional methods. All operations were performed in accordance with the standard operating procedures for cell culture of Beijing Aisiyipu Biotechnology Co., Ltd.

[0598] (3) Determination of the inhibitory effect of compounds on CB1 receptors by homogeneous time-resolved fluorescence (HTRF) assay

[0599] use The cAMP assay was performed using the Ultra cAMP kit (purchased from Perkin Elmer, later renamed Revvity) according to the manufacturer's instructions to determine and compare the inhibitory effects of the compounds on the CB1 receptor.

[0600] Cell treatment: CHO cells overexpressing hCB1 were cultured to 80% confluency, harvested after trypsinization, resuspended in 1× Stimulation Buffer prepared according to the kit instructions, and counted. After dilution, 5 μL of the cell suspension (2500 cells / well) was inoculated into a 384-well plate (purchased from Greiner).

[0601] Preparation of test compounds: The compounds were serially diluted to 10 concentrations and then diluted to 10× with 1× Stimulation Buffer. The final test concentrations of the test compounds were 1000.00 nM, 250.00 nM, 62.50 nM, 15.63 nM, 3.91 nM, 0.98 nM, 0.24 nM, 0.061 nM, 0.015 nM, and 0.0038 nM.

[0602] Experimental steps: 1 μL of diluted compound was added to the corresponding experimental wells and incubated at 37°C for 10 minutes. Then, 4 μL of Forskolin buffer (concentration 2.5 μM, purchased from Selleck) and CP55940 agonist (concentration 2.5 nM, purchased from Sigma) prepared with 1× Stimulation Buffer was added to each well and incubated at 37°C for 30 minutes to induce cAMP production. After the incubation, Eu-cAMP and ULight diluted to the working concentration were added to each well. TM-anti-cAMP (5 μL / well each), centrifuge and incubate at room temperature in the dark for 1 hour. After incubation, use an HTS high-throughput drug screening multifunctional microplate reader (BMG) to obtain the readings at 620 nM and 665 nM of emission light under 330 nm of excitation light.

[0603] GraphPad Prism software was used for nonlinear regression fitting (dose response-variable slope) and the IC of each compound was calculated according to the following equation: 50 : Y=Bottom+(Top-Bottom) / (1+10^((Log IC 50 -X)*HillSlope))

[0604] Wherein, X is the concentration of the test compound, Y is the inhibition rate (%) at the concentration of X, Bottom and Top are the baseline and maximum response, respectively (the unit is the same as Y), and HillSlope is the slope factor.

[0605] At the same time, the CB1 inhibition rate (%) of each compound was calculated using 5 μM rimonabant as a reference according to the following formula:

[0606] Each R value in the formula is the ratio value given by the microplate reader. is the mean of the ratios of all positive control wells (positive control is rimonabant, concentration is 5 μM); The ratios are the mean of all negative control wells (0.1% DMSO).

[0607] (4) Experimental results and analysis

[0608] The results are shown in Figures 1a-1d, 2a-2f, 3a-3e and 4a-4d, which show that positive control compound 1 and positive control compound 2 have significant inhibitory effects on the cAMP upregulation induced by CB1 agonist CP55940, with IC 50 The IC values ​​of compounds Q1-P2, Q2-P2, Q3-P2, Q4-P1, Q5-P1, Q6-P1, Q7-P1, Q8-P2, Q9-P1, Q10-P1, Q11-P1, Q12-P1, Q13-P1, Q14-P2, Q15-P2, Q16-P1, and Q17-P2 were 2.613 nM and 0.2246 nM, respectively. 50The IC values ​​of these compounds are: 0.3507nM, 0.2715nM, 0.3738nM, 0.05826nM, 0.3713nM, 0.4451nM, 0.2458nM, 0.5002nM, 0.4077nM, 0.728nM, 1.493nM, 3.923nM, 0.5142nM, 0.3268nM, 0.09794nM, 0.4359nM and 0.26nM respectively. 50 The activity of the compound of the present invention is comparable to or better than that of the positive control compound, indicating that the compound of the present invention can effectively inhibit CB1.

[0609] Example 2. In vitro CB2 receptor inhibitory activity detection experiment based on HTRF-cAMP assay

[0610] The inhibitory effects of the compounds of this invention on CB2 receptors were analyzed by measuring changes in intracellular cAMP levels using the HTRF (homogeneous time-resolved fluorescence)-cAMP assay. This study employed a CHO cell line stably expressing CB2 receptors. After induction with forskolin (MCE) and CP55940 (Sigma), the inhibitory effects of the compounds on CB2 receptors were determined using an HTRF cAMP assay.

[0611] (1) Test compound and control compound

[0612] In this example, the compounds of the present invention tested include Q1-P2, Q2-P2, Q3-P2, Q4-P1, Q5-P1, Q6-P1, Q7-P1, Q8-P2, Q10-P1, Q11-P1, Q13-P1, Q15-P2, Q16-P1 and Q17-P2, with SR144528 (a known selective CB2 receptor antagonist purchased from MCE) and the control compound 2 in Example 1 as controls.

[0613] (2) Cell construction, culture and passage

[0614] Construction of a CHO cell line stably expressing CB2 receptors: A CHO stable cell line overexpressing human CB2 was constructed using the pCDNA5 / FRT / TO system. Flp-In-CHO cells (CHO cell line purchased from ATCC) were plated at 6×10 5 Cells were seeded into 6-well plates and incubated overnight with complete medium (F12 + 10% FBS) at 37°C and 5% CO2. TMFollowing the instructions for the 3000 transfection kit (purchased from Thermo Fisher), a total of 4 μg of a plasmid containing the human CB2 gene (gene information: NM_001841) was transfected into Flp-In-CHO cells. After 6 hours, the medium was changed. The transfected cells were screened for resistance and then treated with hygromycin for 2-3 weeks to obtain a cell pool stably expressing human CB2. Gene expression was confirmed in the cell line by PCR. Functional analysis (HTRF cAMP assay) was performed on the human CB2-CHO cells after expression was confirmed, resulting in a stable cell line suitable for compound screening.

[0615] Cell culture and passage: The experimental steps, experimental materials and experimental conditions for cell culture and passage were the same as those in Example 1, except that a CHO cell line stably expressing CB2 receptors was used instead of a CHO cell line stably expressing CB1 receptors.

[0616] (3) Determination of the inhibitory effect of compounds on CB2 receptors by homogeneous time-resolved fluorescence (HTRF) experiment

[0617] use cAMP was measured using an Ultra cAMP kit (purchased from Perkin Elmer, later renamed Revvity) according to the manufacturer's instructions to determine and compare the inhibitory effects of the compounds on CB2 receptors. The experimental procedures used the same instruments and conditions as in Example 1, differing only in the use of CHO cells.

[0618] (4) Experimental results and analysis

[0619] The results showed that except for SR144528 as a CB2 inhibitor, the compounds of the present invention and the reference compound 2 had no significant inhibitory effect on CB2: IC 50 The lowest values ​​were 453.8 nM (Q17-P2), 672.1 nM (Q15-P2), 689.6 nM (Q8-P2) and 731.4 nM (Q7-P1). The IC values ​​of other compounds were 50 All of them exceeded 1000 nM, which was much higher than the IC values ​​of the corresponding compounds measured in Example 1 involving CB1 inhibition experiments. 50 The IC value of the control compound SR144528 was 0. 50 It is 67.88nM.

[0620] Example 3. Evaluation of the effect of compounds on inhibiting the binding of CB1 to β-Arrestin2 by NanoBiT method

[0621] NanoBiT is a structurally complementary reporter system that can be used for intracellular detection of protein interactions. It has been applied to a range of research areas, including drug screening, signal transduction analysis, and analysis of viral infection mechanisms. In this experiment, the NanoBiT assay was used to evaluate the effectiveness of compounds in inhibiting the binding of CB1 to β-Arrestin2, a downstream signaling molecule of the cannabinoid receptor CB1.

[0622] (1) Test compound and control compound

[0623] In this example, the compounds of the present invention tested include Q1-P2, Q2-P2, Q3-P2, Q4-P1, Q6-P1, Q7-P1, Q8-P2, Q9-P1, Q10-P1, Q11-P1, Q13-P1, Q15-P2, Q16-P1 and Q17-P2, with the control compound 2 in Example 1 as a control.

[0624] (2) Construction of HEK293T-Smbit-β-Arrestin2 cells stably expressing Smbit-β-Arrestin2

[0625] Using a lentiviral packaging system, the expression vector containing the Smbit-β-Arrestin2 sequence was infected with HEK293T cells via lentivirus to construct HEK293T-Smbit-β-Arrestin2 cells stably expressing Smbit-β-Arrestin2. The specific steps are as follows.

[0626] On the first day, a suspension of HEK-293T cells (purchased from ATCC) was seeded onto a 6 cm culture dish (2 × 10 6 Cells / dish) were cultured at 37°C and 5% CO2 overnight in 5 ml complete medium (DMEM+10% FBS, DMEM purchased from Hyclone, FBS purchased from Avantor).

[0627] On the second day, lentivirus was packaged, specifically by adding Lipofectamine TM 3000 transfection kit (purchased from Thermo Fisher) and Opti-MEM (purchased from Gibco) were placed at room temperature, and then the following system was prepared: Solution A: 100 μl optiMEM + 8 μl Lip3000; Solution B: 100 μl optiMEM + 4 μg Ftasmid + 8 μl P3000, where Lip3000 and P3000 are derived from Lipofectamine TM3000 kit; Ftasmid: PSPAX2 (vector): PVSVG (packaging): plasmid containing Smbit and β-Arrestin2 (2:1:1). Mix Solution A and Solution B and let stand for 15 minutes. Replace the medium in the six-well plate (2 ml of medium per well). After 15 minutes, add the mixed solution to the corresponding six-well plate and mark it. Then, incubate the cells at 37°C and 5% CO2 for 48 hours.

[0628] On the fourth day, the viral supernatant was collected, centrifuged at 1000 rpm for 5 min, and then filtered through a 0.2 μM filter membrane to remove cell particles and debris before being used to infect cells.

[0629] HEK293T cells were plated into 6-well plates at 6 × 10 5 Cells were plated per 6-well plate and incubated with 2 ml of complete culture medium (F12 + 10% FBS) overnight at 37°C and 5% CO2. After replacing the medium in the 6-well plate, 2 ml of viral supernatant was added to each well. 2 μl of polybrene (stock solution of 10 mg / ml) was added to the viral supernatant to a final concentration of 10 μg / ml, mixed thoroughly, and incubated at 37°C and 5% CO2 for 72 hours. After 72 hours, the cells were transferred to different plates and selected for resistance to puromycin to obtain a stably expressing cell line, designated HEK293T-Smbit-β-Arrestin2 cells.

[0630] (3) Construction of HEK293T-Smbit-β-Arrestin2-CB1-Lgbit cells and use of these cells to detect the effect of compounds inhibiting the binding of CB1 to β-Arrestin2

[0631] The HEK293T-Smbit-β-Arrestin2 cells constructed in step (2) were seeded at 700,000 / well in a 6-well plate containing DMEM + 10% FBS (DMEM purchased from Hyclone, FBS purchased from Avantor) and cultured overnight at 37°C. When the cell density reached about 50%, the cells were transfected with Lipofectamine TM The CB1-Lgbit plasmid (obtained by inserting the CB1 gene and Lgbit gene into the pBiT series vector with catalog number N2014 purchased from Promega) was transfected into HEK293T-Smbit-β-Arrestin2 cells using a 3000 transfection kit and cultured at 37°C for 16-24 hours. The resulting HEK293T-Smbit-β-Arrestin2-CB1-Lgbit cells were seeded at 30,000 / well / 70 μL in a 96-well plate purchased from Corning and Opti-MEM TM + 4% FBS (Opti-MEMTM Purchased from Gibco, FBS purchased from Avantor), cultured overnight at 37°C. Cultured cells until confluency reached about 80%, then added 10 μL of 10× diluted antagonist compound (prepared by mixing the compound of the present invention or control compound with DMSO at an initial concentration of 100 nM and diluting 5 times) to the wells, and incubated at 37°C for 12 minutes. Living cell substrates ( Live Cell Substrates, catalog number N2012, purchased from Promega) was diluted to 20 times, 20 μL was added to each well, shaken and set aside. 10 μL of the diluted 10× agonist compound CP55940 (final concentration 100 nM) was added to the wells, gently shaken and incubated in the dark for 10 minutes. After the incubation, the cells were analyzed using a microplate reader ( FSX, purchased from BMG) was used to detect luminescence and record the fluorescence value. Nonlinear regression fitting (dose response-variable slope) was performed using GraphPad Prism software, and the IC of each compound was calculated according to the following equation: 50 : Y=Bottom+(Top-Bottom) / (1+10^((Log IC 50 -X)*HillSlope))

[0632] Wherein, X is the concentration of the test compound, Y is the inhibition rate (%) at the concentration of X, Bottom and Top are the baseline and maximum response, respectively (the unit is the same as Y), and HillSlope is the slope factor.

[0633] The CB1 inhibition rate (%) of each compound was calculated using 5 μM rimonabant as a reference according to the following formula: Inhibition rate (%) = (Sample-Ave_H) / (Ave_L-Ave_H)*100

[0634] The values ​​in the formula are the fluorescence values ​​given by the microplate reader, Ave_L is the average value of all positive control wells (positive control is rimonabant, the concentration is 5 μM); Ave_H is the average value of all negative control wells (0.1% DMSO).

[0635] (4) Experimental results and analysis

[0636] The results showed that the compounds Q1-P2, Q2-P2, Q3-P2, Q4-P1, Q6-P1, Q7-P1, Q8-P2, Q9-P1, Q10-P1, Q11-P1, Q13-P1, Q15-P2, Q16-P1 and Q17-P2 of the present invention can effectively inhibit the binding of CB1 to β-Arrestin2, and their IC 50The inhibitory effects were as follows: 0.2757nM, 0.393nM, 1.292nM, 0.326nM, 0.043nM, 0.09nM, 0.274nM, 0.726nM, 0.21nM, 0.541nM, 0.318nM, 0.282nM, 0.1408nM and 1.092nM, respectively. 50 was 0.703 nM) was comparable or even better.

[0637] Example 4. Pharmacokinetic (PK) evaluation

[0638] The pharmacokinetic (PK) evaluation of the compounds of the present invention was performed using 6-8 week old ICR male mice weighing approximately 25 g purchased from Jiangsu Huachuang Xinnuo Pharmaceutical Technology Co., Ltd. All mice were housed in IVC (independent ventilation system) cages in an SPF animal room. The animal information card indicated the number of animals in each cage, sex, strain, receipt date, dosing regimen, experiment number, group, and start date of the experiment. All cages, bedding, and drinking water were sterilized before use. The experimental animals were fasted for more than 12 hours during the experiment, had free access to drinking water, and were provided with food 4 hours after administration. The experimental animals were identified with ear tags.

[0639] Using 0.5% HPMC-E5 (purchased from Shanghai Saiyi Biotechnology Co., Ltd., 0.5 g of HPMC-E5 was diluted to 100 mL of aqueous solution to prepare a 0.5% solution) as the solvent, the test compound or control compound was prepared into a suspension (the dosing suspension concentration was 1.0 mg / mL). Mice were divided into groups, with 21 mice per group. The mice were orally administered with the compound of the present invention or the control compound by gavage at a dose of 10 mg / kg in a dosing volume of 10 ml / kg.

[0640] The animals (3 mice per group at each time point) were anesthetized at 0.25, 0.5, 1, 2, 4, 8 and 24 hours after administration, and sacrificed by exsanguination via the abdominal aorta. Approximately 400 microliters of blood was collected through the appropriate vein, placed in an EDTA-K2 test tube, and centrifuged at 11000 rpm for 5 minutes to separate the plasma. All brain tissues were dissected and collected, rinsed with ice saline and blotted dry with filter paper. All plasma and brain tissues were placed in a -70°C refrigerator and frozen for testing. The concentrations of each drug in plasma and brain tissue were determined by LC-MC / MS, and the following parameters were obtained by Phoenix WinNonlin analysis. The results are shown in Table 1 below.

[0641] Table 1 Average pharmacokinetic parameters after single oral administration in mice

[0642] MRT0-∞ : The average residence time of the drug in the whole body;

[0643] AUC 0-t : The area under the drug concentration-time curve from time 0 to the sample collection time t at which the last quantifiable concentration can be obtained;

[0644] AUC 0-∞ : The area under the drug concentration-time curve from time 0 to infinite time;

[0645] C max : Maximum observed plasma concentration;

[0646] T max : time of maximum observed plasma concentration;

[0647] t 1 / 2 : Apparent plasma terminal elimination half-life.

[0648] The above results show that compared with the plasma exposure, the brain tissue exposure of the tested compounds is very low or below the detection limit, which indicates that the compounds hardly enter the brain or have extremely low brain penetration in mice, and have high peripheral selectivity.

[0649] Industrial Applicability

[0650] The present invention provides a CB1 inhibitor that can be used to treat or prevent obesity or obesity complications. Therefore, the present invention is suitable for industrial application.

[0651] Although the present invention is described in detail herein, the present invention is not limited thereto. Those skilled in the art may make modifications based on the principles of the present invention. Therefore, all modifications made in accordance with the principles of the present invention should be understood to fall within the scope of protection of the present invention.

Claims

1. A compound of formula (I), or a pharmaceutically acceptable salt, cocrystal, stereoisomer, solvate, prodrug or metabolite thereof: in, Ring A represents a five-membered or six-membered monocyclic heteroaryl group containing 1 or 2 heteroatoms independently selected from N and S; R1 is C 1-6 alkyl halide; m is 0, 1 or 2, R2 is independently selected from halogen, cyano, and C 1-6 alkyl halide; R 3a 、R 3b 、R 4a and R 4b are each independently selected from hydrogen and C 1-6 alkyl.

2. The compound of formula (I) according to claim 1, or a pharmaceutically acceptable salt, cocrystal, stereoisomer, solvate, prodrug or metabolite thereof, wherein: Ring A is selected from thienyl, pyridyl and thiazolyl; R1 is C 1-3 alkyl halide; m is 1 or 2, R2 is independently selected from fluorine, chlorine, cyano, and C 1-3 alkyl halide; R 3a 、R 3b 、R 4a and R 4b are each independently selected from hydrogen and C 1-3 alkyl.

3. The compound of formula (I) according to claim 2, or a pharmaceutically acceptable salt, cocrystal, stereoisomer, solvate, prodrug or metabolite thereof, wherein: Ring A is selected from thien-2-yl, pyridin-2-yl and thiazol-5-yl; R1 is C 1-3 Fluorinated alkyl; m is 1, R2 is C 1-3 Fluorinated alkyl; R 3a and R 3b At least one of them is hydrogen, and R 4a and R 4b Each is hydrogen.

4. A compound of formula (I) according to any one of claims 1 to 3, or a pharmaceutically acceptable salt, cocrystal, stereoisomer, solvate, prodrug or metabolite thereof, wherein the compound is selected from:

5. A compound of formula (II), or a pharmaceutically acceptable salt, cocrystal, stereoisomer, solvate, prodrug or metabolite thereof: in, Ring B represents a phenyl group or a five-membered or six-membered monocyclic heteroaryl group containing 1 or 2 heteroatoms independently selected from N and S; Ring C represents a phenyl group or a thienyl group; R1 is C 1-6 alkyl halide; m is 0, 1 or 2, R2 is independently selected from halogen, cyano, and C 1-6 alkyl halide; R 3a 、R 3b 、R 4a and R 4b are each independently selected from hydrogen and C 1-6 alkyl.

6. The compound of formula (II) according to claim 5, or a pharmaceutically acceptable salt, cocrystal, stereoisomer, solvate, prodrug or metabolite thereof, wherein Ring B is selected from phenyl, thienyl, and pyridyl; Ring C is selected from phenyl and thiophen-2-yl; R1 is C 1-3 alkyl halide; m is 1 or 2, R2 is independently selected from fluorine, chlorine, cyano, and C 1-3 alkyl halide; R 3a 、R 3b 、R 4a and R 4b are each independently selected from hydrogen and C 1-3 alkyl.

7. The compound of formula (II) according to claim 6, or a pharmaceutically acceptable salt, cocrystal, stereoisomer, solvate, prodrug or metabolite thereof, wherein Ring B is selected from phenyl, thien-2-yl, and pyridin-2-yl; Ring C is selected from phenyl; R1 is C 1-3 Fluorinated alkyl; m is 1, R2 is C 1-3 Fluorinated alkyl; R 3a and R 3b At least one of them is hydrogen, and R 4a and R 4b Each is hydrogen.

8. A compound of formula (II) according to any one of claims 5 to 7, or a pharmaceutically acceptable salt, cocrystal, stereoisomer, solvate, prodrug or metabolite thereof, wherein the compound is selected from:

9. A compound, or a pharmaceutically acceptable salt, cocrystal, stereoisomer, solvate, prodrug or metabolite thereof, wherein the compound is selected from:

10. A pharmaceutical composition comprising: (1) The compound according to any one of claims 1 to 9, or a pharmaceutically acceptable salt, cocrystal, stereoisomer, solvate, prodrug or metabolite thereof; (2) one or more other active ingredients; as well as (3) Pharmaceutically acceptable carriers and / or excipients.

11. Use of the compound according to any one of claims 1 to 9, or a pharmaceutically acceptable salt, cocrystal, stereoisomer, solvate, prodrug or metabolite thereof, in the preparation of a medicament for treating or preventing obesity or obesity complications.

12. The use according to claim 11, wherein the obesity complications are selected from one or more of diabetes, dyslipidemia, metabolic syndrome, dementia, inflammatory disease, cardiovascular disease, liver disease, or cancer.

13. The use according to claim 12, wherein The diabetes is selected from one or more of type 1 diabetes, type 2 diabetes, impaired glucose tolerance or insulin resistance; The dyslipidemia is selected from one or more of poor blood lipid levels, low levels of high-density lipoprotein, high levels of low-density lipoprotein or high levels of triglycerides; The inflammatory disease is selected from one or more of osteoarthritis, rheumatoid arthritis, inflammatory bowel disease or obesity-related inflammation; The cardiovascular disease is selected from one or more of atherosclerosis, hypertension, stroke or heart attack; The liver disease is selected from one or more of liver inflammation, liver fibrosis, non-alcoholic steatohepatitis, fatty liver, liver enlargement, alcoholic liver disease, jaundice, cirrhosis or hepatitis; The cancer is selected from one or more of colon cancer, breast cancer, thyroid cancer, alveolar rhabdomyosarcoma or hepatocellular carcinoma.

14. The method of claim 11, wherein the obesity complication is selected from one or more of hypertension, gallbladder disease, gastrointestinal disease, menstrual irregularities, degenerative arthritis, venous stasis ulcers, pulmonary hypoventilation syndrome, sleep apnea, snoring, coronary artery disease, arteriosclerotic disease, pseudotumor cerebri, accident proneness, increased surgical risk, osteoarthritis, high cholesterol, or an increased incidence of ovarian, cervical, uterine, breast, prostate, or gallbladder malignancies.

15. Use of the compound according to any one of claims 1 to 9, or a pharmaceutically acceptable salt, cocrystal, stereoisomer, solvate, prodrug or metabolite thereof, in a medicament for treating or preventing obesity or complications of obesity.

16. Use of the compound of claim 15, or a pharmaceutically acceptable salt, cocrystal, stereoisomer, solvate, prodrug or metabolite thereof, wherein the obesity complication is selected from one or more of diabetes, dyslipidemia, metabolic syndrome, dementia, inflammatory disease, cardiovascular disease, liver disease, or cancer.

17. The use of the compound of claim 16, or a pharmaceutically acceptable salt, cocrystal, stereoisomer, solvate, prodrug or metabolite thereof, wherein: The diabetes is selected from one or more of type 1 diabetes, type 2 diabetes, impaired glucose tolerance or insulin resistance; The dyslipidemia is selected from one or more of poor blood lipid levels, low levels of high-density lipoprotein, high levels of low-density lipoprotein or high levels of triglycerides; The inflammatory disease is selected from one or more of osteoarthritis, rheumatoid arthritis, inflammatory bowel disease or obesity-related inflammation; The cardiovascular disease is selected from one or more of atherosclerosis, hypertension, stroke or heart attack; The liver disease is selected from one or more of liver inflammation, liver fibrosis, non-alcoholic steatohepatitis, fatty liver, liver enlargement, alcoholic liver disease, jaundice, cirrhosis or hepatitis; The cancer is selected from one or more of colon cancer, breast cancer, thyroid cancer, alveolar rhabdomyosarcoma or hepatocellular carcinoma.

18. The use of a compound according to claim 15, or a pharmaceutically acceptable salt, cocrystal, stereoisomer, solvate, prodrug or metabolite thereof, wherein the obesity complication is selected from one or more of hypertension; gallbladder disease; gastrointestinal disease; menstrual irregularities; degenerative arthritis; venous stasis ulcers; pulmonary hypoventilation syndrome; sleep apnea; snoring; coronary artery disease; arteriosclerotic disease; pseudotumor cerebri; accident proneness; increased surgical risk; osteoarthritis; high cholesterol; or an increased incidence of ovarian, cervical, uterine, breast, prostate or gallbladder malignancies.

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