Inhibitors of human immunodeficiency virus replication

Abstract

To provide a pharmaceutical composition useful for treating HIV infection. [Solution] A pharmaceutical composition is provided that includes, for example, the following compound or a pharmaceutically acceptable salt thereof. JPEG2026062839000055.jpg62170

Description

[Technical Field]

[0001] The present invention relates to pharmaceutical compositions and methods for using these compositions in the treatment of HIV infection. [Background technology]

[0002] Acquired immunodeficiency syndrome (AIDS) is a result of HIV infection. HIV remains a major global public health problem. In 2015, an estimated 36.7 million people were living with HIV (including 1.8 million children), and the global HIV prevalence was 0.8%. The majority of these people live in low- and middle-income countries. In the same year, 1.1 million people died from AIDS-related illnesses.

[0003] Current therapies for HIV-infected individuals consist of combinations of approved antiretroviral drugs. Currently, nearly four dozen drugs are approved for HIV infection, either as monotherapy, fixed-dose combinations, or single-tablet regimens (the latter two including 2-4 approved drugs). These drugs belong to several different classes, targeting either viral enzymes or the function of viral proteins during the viral replication cycle. Therefore, drugs are classified as either nucleoside reverse transcriptase inhibitors (NRTIs), non-nucleoside reverse transcriptase inhibitors (NNRTIs), protease inhibitors (PIs), integrase chain transfer inhibitors (INSTIs), or entry inhibitors (one of which is maraviroc, targeting the host CCR5 protein, while another is enfvirtide, a peptide targeting the gp41 region of the viral gp160 protein). In addition, pharmacokinetic enhancers (cobicistat or ritonavir) can be used in combination with antiretroviral drugs (ARVs) that require a boost.

[0004] Despite the availability of medical equipment consisting of single-agent and combination drugs, the medical need for new antiretroviral drugs remains. High viral heterogeneity, drug-related toxicity, tolerability issues, and poor adhesion can all lead to treatment failure and result in the selection of viruses with mutations that confer resistance to one or more antiretroviral agents or even more drugs from the entire class (Beyrer, C., Pozniak A. HIV drug resistance - an emerging threat to epidemic control. N. Engl. J. Med. 2017, 377, 1605-1607; Gupta, RK, Gregson J., et al. HIV-1 drug resistance before initiation or re-initiation of first-line antiretroviral therapy in low-income and middle-income countries: a systematic review and meta-regression analysis. Lancet Infect. Dis. 2017, 18, 346-355; Zazzi, M., Hu, H., Prosperi, M. The global burden of HIV-1 drug resistance in the past 20 years. (PeerJ. 2018, DOI 10.7717 / peerj.4848). As a result, there is a need for new drugs that are easier to administer, have a high genetic barrier to the development of resistance, and are safer than current drugs. Among these numerous options, novel mechanisms of action (MOAs) that can be used as part of preferred antiretroviral therapy (ART) may still have a major role to play, as they should be effective against viruses resistant to current drugs.

[0005] Compounds with certain therapeutic potential are currently described in the art, as described in Blair, Wade S. et.al. Antimicrobial Agents and Chemotherapy (2009), 53(12), 5080-5087, Blair, Wade S. et al. PLoS Pathogens (2010), 6(12), e1001220, Thenin-Houssier, Suzie; Valente, Susana T. Current HIV Research, 2016, 14, 270-282 and the following numbered PCT patent applications: WO2012065062, WO2013006738, WO2013006792, WO2014110296, WO2014110297, WO2014110298, WO2014134566, WO2015130964, WO2015130966, WO2016033243, WO2018035359, WO2018203235, WO2019161017 and WO2019161280.

[0006] Compounds currently needed in the art are new and additional compounds useful in the treatment of HIV. In addition, these compounds should offer advantages for pharmaceutical use, for example, with respect to one or more of their mechanism of action, binding, inhibitory potency, target selectivity, solubility, safety profile, bioavailability and / or reduction in dosing frequency. New formulations and treatment methods using these compounds are also needed. Formulations of certain compounds are disclosed in WO2020 / 018459. SUMMARY OF THE INVENTION

[0007] Briefly, in one aspect, the present invention relates to formula Ia, formula Ib, formula Ic or formula Id:

Chemical formula

[0008] In another aspect, the present invention discloses a method for treating HIV infection in humans, which comprises administering the compound or salt of the present invention.

[0009] In another aspect, the present invention discloses the compound or salt of the present invention for use in therapy.

[0010] In another aspect, the present invention discloses the compound or salt of the present invention for use in the treatment of HIV infection in humans.

[0011] In another aspect, the present invention discloses the use of the compound or salt of the present invention in the manufacture of a medicament for the treatment of HIV infection in humans.

Brief Description of the Drawings

[0012] [Figure 1] Figure 1 is a diagram showing the outline of the PK experiment described below.

Modes for Carrying Out the Invention

[0013] In one aspect, the pharmaceutical composition of the present invention further comprises an additive selected from the group consisting of polysorbate 20, polysorbate 80, poloxamer 188, poloxamer 207, poloxamer 338, sodium chloride, and sodium hydroxide.

[0014] In one aspect, the pharmaceutical composition of the present invention is a solution.

[0015] In one aspect, the pharmaceutical composition of the present invention is a suspension.

[0016] In one embodiment, the pharmaceutical composition of the present invention contains water.

[0017] In one embodiment, the pharmaceutical composition of the present invention comprises polyethylene glycol (PEG).

[0018] In one embodiment, the pharmaceutical composition of the present invention contains polyethylene glycol (PEG), the average molecular weight of PEG being approximately 300 (PEG300).

[0019] In one embodiment, the pharmaceutical composition of the present invention contains ethanol.

[0020] In one embodiment, the pharmaceutical composition of the present invention contains ethyl lactate.

[0021] In one embodiment, the pharmaceutical composition of the present invention comprises poloxamer 188.

[0022] In one embodiment, the pharmaceutical composition of the present invention contains sodium hydroxide.

[0023] In one embodiment, the pharmaceutical composition of the present invention is a pharmaceutical composition in which at least 90% by weight of the solvent or diluent is water and PEG300.

[0024] In one embodiment, the pharmaceutical composition of the present invention is a pharmaceutical composition in which at least 90% by weight of the additive is poloxamer 188.

[0025] In one embodiment, the pharmaceutical composition of the present invention is a pharmaceutical composition in which water is present in an amount of 8 to 20% by weight and PEG300 is present in an amount of 60 to 85% by weight.

[0026] In one embodiment, the pharmaceutical composition of the present invention is a pharmaceutical composition in which water is present in an amount of 8 to 12% by weight and PEG300 is present in an amount of 63 to 70% by weight.

[0027] In one embodiment, the pharmaceutical composition of the present invention is a pharmaceutical composition in which a compound of formula Ia, formula Ib, formula Ic, or formula Id, or a pharmaceutically acceptable salt thereof, is present in the composition at a concentration of 50 to 500 mg / mL by weight of the free compound of formula Ia, formula Ib, formula Ic, or formula Id. Throughout this specification, references to concentrations of compounds of formula Ia, Ib, Ic, or Id, or their pharmaceutically acceptable salts, always refer to concentrations based on the free compounds of formula Ia, Ib, Ic, or Id, and not concentrations based on their salts.

[0028] In one embodiment, the pharmaceutical composition of the present invention is a pharmaceutical composition in which a compound of formula Ia, formula Ib, formula Ic, or formula Id, or a pharmaceutically acceptable salt thereof, is present in the composition at a concentration of 225 to 275 mg / mL.

[0029] In one embodiment, the pharmaceutical composition of the present invention is a pharmaceutical composition in which a compound of formula Ia, formula Ib, formula Ic, or formula Id, or a pharmaceutically acceptable salt thereof, is present in the composition at a concentration of 275 to 350 mg / mL.

[0030] In one embodiment, the pharmaceutical composition of the present invention is a pharmaceutical composition in which a compound of formula Ia, formula Ib, formula Ic, or formula Id, or a pharmaceutically acceptable salt thereof, is present in the composition at a concentration of 350 to 425 mg / mL.

[0031] In one embodiment, the pharmaceutical composition of the present invention is prepared from the crystalline form of a compound of formula Ia, formula Ib, formula Ic, or formula Id, or a pharmaceutically acceptable salt thereof. In another embodiment, the pharmaceutical composition of the present invention is a suspension of the crystalline form of a compound of formula Ia, formula Ib, formula Ic, or formula Id, or a pharmaceutically acceptable salt thereof.

[0032] In one embodiment, the pharmaceutical composition of the present invention is prepared from an amorphous form of a compound of formula Ia, formula Ib, formula Ic, or formula Id, or a pharmaceutically acceptable salt thereof. In another embodiment, the amorphous form is a lyophilized powder. In yet another embodiment, the pharmaceutical composition of the present invention is a suspension of the amorphous form.

[0033] In another embodiment, the formulation contains water. In another embodiment, the formulation contains glycoflor. In another embodiment, the formulation contains N-methyl-2-pyrrolidone (NMP). In another embodiment, the formulation contains dimethyl sulfoxide (DMSO). In another embodiment, the formulation contains alcohol. In another embodiment, the formulation contains ethanol. In another embodiment, the formulation contains physiological saline. In another embodiment, the formulation contains polyethylene glycol (PEG), whose chemical formula is generally H-(O-CH2-CH2) n It can be represented as -OH. In another embodiment, the formulation contains PEG (PEG100) with an average molecular weight of about 100. In another embodiment, the formulation contains PEG (PEG200) with an average molecular weight of about 200. In another embodiment, the formulation contains PEG (PEG300) with an average molecular weight of about 300. In another embodiment, the formulation contains PEG (PEG400) with an average molecular weight of about 400. In another embodiment, the formulation contains PEG (PEG500) with an average molecular weight of about 500. In another embodiment, the formulation contains PEG (PEG600) with an average molecular weight of about 600. In another embodiment, the formulation contains PEG "capped" by an alkyl group, and as a result, the formula of PEG is generally alkyl-(O-CH2-CH2) n It may be represented as -O-alkyl. In another embodiment, the component constituting more than 50 w / w% of the formulation is other than water. The component constituting more than 60 w / w% of the formulation is other than water. The component constituting more than 70 w / w% of the formulation is other than water. The component constituting more than 80 w / w% of the formulation is other than water. The component constituting more than 85 w / w% of the formulation is other than water. The component constituting more than 90 w / w% of the formulation is other than water. In another embodiment, the formulation contains water and PEG200. In another embodiment, the formulation contains water and PEG300.

[0034] In one embodiment, the pharmaceutical composition of the present invention comprises a compound of formula Ia, formula Ib, formula Ic, or formula Id, or a pharmaceutically acceptable salt thereof, and a solubilizing additive. In another embodiment, the solubilizing additive is polysorbate. In another embodiment, the solubilizing additive is polysorbate 20. In another embodiment, the solubilizing additive is polysorbate 80. In another embodiment, the solubilizing additive is poloxamer. In another embodiment, the solubilizing additive is poloxamer 188 (P188). In another embodiment, the solubilizing additive is poloxamer 207. In another embodiment, the solubilizing additive is poloxamer 338 (P338). In another embodiment, the solubilizing additive is docusate sodium. In another embodiment, the solubilizing additive is polyethylene glycol 3350 (PEG3350). In another embodiment, the solubilizing additive is a fatty acid. In another embodiment, the solubilizing additive is a fatty acid containing 8 to 12 carbon atoms. In another embodiment, the solubilizing additive is a fatty acid containing eight carbon atoms (C8). In another embodiment, the solubilizing additive is a fatty acid containing twelve carbon atoms (C12). In another embodiment, the solubilizing additive is less than 5 w / w% of the formulation. In another embodiment, the solubilizing additive is less than approximately 5 w / w% of the formulation. In another embodiment, the solubilizing additive is less than approximately 4 w / w% of the formulation. In another embodiment, the solubilizing additive is less than approximately 3 w / w% of the formulation. In another embodiment, the solubilizing additive is less than approximately 2 w / w% of the formulation. In another embodiment, the solubilizing additive is less than approximately 1 w / w% of the formulation. In another embodiment, the solubilizing additive is less than approximately 0.5 w / w% of the formulation. In another embodiment, the solubilizing additive is less than approximately 0.25 w / w% of the formulation.

[0035] In one embodiment, the pharmaceutical composition of the present invention comprises poloxamer 188 and C8 fatty acid. In another embodiment, the ratio of poloxamer 188 to C8 fatty acid is approximately 3:1. In yet another embodiment, the concentration of poloxamer 188 is approximately 3 w / w% of the formulation, and the concentration of C8 fatty acid is approximately 1 w / w% of the formulation.

[0036] In one embodiment, the pharmaceutical composition of the present invention comprises poloxamer 188 and C12 fatty acid. In another embodiment, the ratio of poloxamer 188 to C12 fatty acid is approximately 6:1. In yet another embodiment, the concentration of poloxamer 188 is approximately 3 w / w% of the formulation, and the concentration of C12 fatty acid is approximately 0.5 w / w% of the formulation.

[0037] In one embodiment, the pharmaceutical composition of the present invention comprises poloxamer 338 and C8 fatty acid. In another embodiment, the ratio of poloxamer 338 to C8 fatty acid is approximately 3:1. In yet another embodiment, the concentration of poloxamer 338 is approximately 3 w / w% of the formulation, and the concentration of C8 fatty acid is approximately 1 w / w% of the formulation.

[0038] In one embodiment, the pharmaceutical composition of the present invention comprises poloxamer 338 and C12 fatty acid. In another embodiment, the ratio of poloxamer 338 to C12 fatty acid is approximately 6:1. In yet another embodiment, the concentration of poloxamer 338 is approximately 3 w / w% of the formulation, and the concentration of C12 fatty acid is approximately 0.5 w / w% of the formulation.

[0039] In one embodiment, the pharmaceutical composition of the present invention comprises docusate sodium and C8 fatty acid. In another embodiment, the ratio of docusate sodium to C8 fatty acid is about 3:1. In yet another embodiment, the concentration of docusate sodium is about 3 w / w% of the formulation, and the concentration of C8 fatty acid is about 1 w / w% of the formulation.

[0040] In one embodiment, the pharmaceutical composition of the present invention comprises docusate sodium and C12 fatty acid. In another embodiment, the ratio of docusate sodium to C12 fatty acid is about 6:1. In yet another embodiment, the concentration of docusate sodium is about 3 w / w% of the formulation, and the concentration of C12 fatty acid is about 0.5 w / w% of the formulation.

[0041] In one embodiment, the pharmaceutical composition of the present invention contains a chelating agent. In another embodiment, the chelating agent is Na-EDTA. In yet another embodiment, Na-EDTA is present in the formulation at a concentration of 0.01 to 0.05 w / w%.

[0042] In one embodiment, the pharmaceutical composition of the present invention contains an antioxidant. In another embodiment, the antioxidant is L-methionine. In another embodiment, the antioxidant is vitamin E. In yet another embodiment, the antioxidant is present in the formulation at a concentration of 0.01 to 0.10 w / w%.

[0043] In one embodiment, the pharmaceutical composition of the present invention contains an acid. In another embodiment, the acid is ethanesulfonic acid. In another embodiment, the concentration of ethanesulfonic acid is 1 to 10 mM. In another embodiment, the concentration of ethanesulfonic acid is 4.8 to 5.2 mM. In another embodiment, the acid is methanesulfonic acid. In another embodiment, the concentration of methanesulfonic acid is 1 to 10 mM. In another embodiment, the concentration of methanesulfonic acid is 4.8 to 5.2 mM.

[0044] In one embodiment, the pharmaceutical composition of the present invention contains a base. In another embodiment, the base is an inorganic base. In another embodiment, the base is an organic base. In another embodiment, the anion of the base is - It is OH (hydroxide). In another embodiment, the anion of a base is - It is OEt (ethoxide). In another embodiment, the anion of a base is - It is OAc (acetate). In another embodiment, the cation of the base is + It is Na (sodium). In another embodiment, the cation of a base is +In another embodiment, the cation of the base is choline. In another embodiment, the base is sodium acetate. In another embodiment, the base is sodium hydroxide. In another embodiment, the base is sodium ethoxide. In another embodiment, the base is choline hydroxide. In another embodiment, the pharmaceutical composition of the present invention contains 0.1 to 1.5 molar equivalents of the base relative to the compound of formula Ia, formula Ib, formula Ic, or formula Id. In another embodiment, the pharmaceutical composition of the present invention contains 0.7 to 1.2 molar equivalents of the base relative to the compound of formula Ia or formula Ib. In another embodiment, the pharmaceutical composition of the present invention contains 0.7 to 1.0 molar equivalents of the base relative to the compound of formula Ia or formula Ib. In another embodiment, the pharmaceutical composition of the present invention contains 1.0 to 1.2 molar equivalents of the base relative to the compound of formula Ia or formula Ib. In another embodiment, the pharmaceutical composition of the present invention contains approximately equimolar equivalents of the base and the compound of formula Ia, formula Ib, formula Ic, or formula Id.

[0045] In one embodiment, the pharmaceutical composition of the present invention comprises a buffer. In another embodiment, the buffer is an acetate buffer. In another embodiment, the pH of the buffer is 4.0 to 7.4. In another embodiment, the pH of the buffer is 4.0 to 5.5. In another embodiment, the pH of the buffer is 4.8 to 5.2. In another embodiment, the concentration of the buffer is 1 to 10 mM. In another embodiment, the concentration of the buffer is 4.8 to 5.2 mM.

[0046] In one embodiment, the pharmaceutical composition of the present invention contains 50 to 500 mg / mL of a compound of formula Ia, formula Ib, formula Ic, or formula Id, or a pharmaceutically acceptable salt thereof. In another embodiment, the pharmaceutical composition of the present invention contains about 50 mg / mL of a compound of formula Ia, formula Ib, formula Ic, or formula Id, or a pharmaceutically acceptable salt thereof. In another embodiment, the pharmaceutical composition of the present invention contains about 100 mg / mL of a compound of formula Ia, formula Ib, formula Ic, or formula Id, or a pharmaceutically acceptable salt thereof. In another embodiment, the pharmaceutical composition of the present invention contains about 150 mg / mL of a compound of formula Ia, formula Ib, formula Ic, or formula Id, or a pharmaceutically acceptable salt thereof. In another embodiment, the pharmaceutical composition of the present invention contains about 200 mg / mL of a compound of formula Ia, formula Ib, formula Ic, or formula Id, or a pharmaceutically acceptable salt thereof. In another embodiment, the pharmaceutical composition of the present invention contains about 250 mg / mL of a compound of formula Ia, formula Ib, formula Ic, or formula Id, or a pharmaceutically acceptable salt thereof. In another embodiment, the pharmaceutical composition of the present invention contains about 300 mg / mL of a compound of formula Ia, formula Ib, formula Ic, or formula Id, or a pharmaceutically acceptable salt thereof. In another embodiment, the pharmaceutical composition of the present invention contains about 350 mg / mL of a compound of formula Ia, formula Ib, formula Ic, or formula Id, or a pharmaceutically acceptable salt thereof. In another embodiment, the pharmaceutical composition of the present invention contains about 400 mg / mL of a compound of formula Ia, formula Ib, formula Ic, or formula Id, or a pharmaceutically acceptable salt thereof. In another embodiment, the pharmaceutical composition of the present invention contains about 450 mg / mL of a compound of formula Ia, formula Ib, formula Ic, or formula Id, or a pharmaceutically acceptable salt thereof. In another embodiment, the pharmaceutical composition of the present invention comprises about 500 mg / mL of a compound of formula Ia, formula Ib, formula Ic, or formula Id, or a pharmaceutically acceptable salt thereof.

[0047] In one embodiment, the pharmaceutical composition of the present invention comprises a compound of formula Ia, formula Ib, formula Ic, or formula Id or a pharmaceutically acceptable salt thereof, water, PEG300, and 0 to 3 additional additives.

[0048] In one embodiment, the pharmaceutical composition of the present invention comprises a compound of formula Ia, formula Ib, formula Ic, or formula Id or a pharmaceutically acceptable salt thereof, water, PEG200, and 0 to 3 additional additives.

[0049] In one embodiment, the pharmaceutical composition of the present invention comprises a compound of formula Ia, formula Ib, formula Ic, or formula Id or a pharmaceutically acceptable salt thereof, poloxamer P338, and acetate buffer. In another embodiment, the concentration of the compound of formula Ia, formula Ib, formula Ic, or formula Id or a pharmaceutically acceptable salt thereof is 280 to 320 mg / mL. In another embodiment, the concentration of poloxamer P338 is 3 to 5 w / w%. In another embodiment, the pH of the acetate buffer is 4.8 to 5.2. In another embodiment, the concentration of the acetate buffer is 4.8 to 5.2 mM.

[0050] In one embodiment, the pharmaceutical composition of the present invention comprises a compound of formula Ia, formula Ib, formula Ic, or formula Id or a pharmaceutically acceptable salt thereof, poloxamer P338, ethanesulfonic acid, and acetate buffer. In another embodiment, the concentration of the compound of formula Ia, formula Ib, formula Ic, or formula Id or a pharmaceutically acceptable salt thereof is 280 to 320 mg / mL. In another embodiment, the concentration of poloxamer P338 is 3 to 5 w / w%. In another embodiment, the concentration of ethanesulfonic acid is 4.8 to 5.2 mM. In another embodiment, the pH of the acetate buffer is 4.8 to 5.2. In another embodiment, the concentration of the acetate buffer is 4.8 to 5.2 mM.

[0051] In one embodiment, the pharmaceutical composition of the present invention comprises a compound of formula Ia, formula Ib, formula Ic, or formula Id or a pharmaceutically acceptable salt thereof, poloxamer P338, C8-C12 fatty acids, and acetate buffer. In another embodiment, the concentration of the compound of formula Ia, formula Ib, formula Ic, or formula Id or a pharmaceutically acceptable salt thereof is 280-320 mg / mL. In another embodiment, the concentration of poloxamer P338 is 3-5 w / w%. In another embodiment, the concentration of fatty acids is 0.5-1 w / w%. In another embodiment, the pH of the acetate buffer is 4.8-5.2. In another embodiment, the concentration of the acetate buffer is 4.8-5.2 mM.

[0052] In one embodiment, the pharmaceutical composition of the present invention comprises a compound of formula Ia, formula Ib, formula Ic, or formula Id or a pharmaceutically acceptable salt thereof, docusate sodium, and acetate buffer. In another embodiment, the concentration of the compound of formula Ia, formula Ib, formula Ic, or formula Id or a pharmaceutically acceptable salt thereof is 280 to 320 mg / mL. In another embodiment, the concentration of docusate sodium is 1 to 2 w / w%. In another embodiment, the pH of the acetate buffer is 4.8 to 5.2. In another embodiment, the concentration of the acetate buffer is 4.8 to 5.2 mM.

[0053] In one embodiment, the present invention discloses a pharmaceutical composition, wherein the pharmaceutical composition is a solution and has been sterilized by passing it through a filter.

[0054] The salts of the present invention are pharmaceutically acceptable. Such salts may be acid addition salts or base addition salts. For a general overview of suitable pharmaceutically acceptable salts, see, for example, Berge et al, J. Pharm, Sci., 66, 1-19, 1977.

[0055] Representative pharmaceutically acceptable acid addition salts include, but are not limited to, 4-acetamidebenzoate, acetate, adipine, alginate, ascorbate, aspartate, benzenesulfonate (besylate), benzoate, bisulfate, tartrate, butyrate, calcium edetate, camphorate, camphor sulfonate (cansylate), caprate (decanoate), caproate (hexanoate), caprylate (octanoate), cinnamate, citrate, cyclamate, digluconate, and 2,5-dihydroxybenzoate. , disuccinate, dodecyl sulfate (estolate), edetate (ethylenediaminetetraacetate), estolate (lauryl sulfate), ethane-1,2-disulfonate (edisylate), ethanesulfonate (esylate), formate, fumarate, galactarate (mucinate), gentisinate (2,5-dihydroxybenzoate), glucoheptoneate (gluceptate), gluconate, glucuronate, glutamate, glutarate, glycerophosphate, glycolate, hexylresorcinate, hippurate, hydravamin (N, N'-di(dehydroabiethyl)-ethylenediamine), hydrobromide, hydrochloride, hydroiodide, hydroxynaphthoate, isobutyrate, lactate, lactobionate, laurate, malate, maleate, malonate, mandelate, methanesulfonate (mesylate), methylsulfate, mucinate, naphthalene-1,5-disulfonate (napadisylate), naphthalene-2-sulfonate (napsylate), nicotinate, nitrate, oleate, palmitate, p-aminobenzenesulfonate, p-aminosalicylate, pa Examples include motates (embonates), pantothenates, pectinates, persulfates, phenylacetates, phenylethylbarbitalates, phosphates, polygalacturonates, propionates, p-toluenesulfonates (tosylates), pyroglutamates, pyruvates, salicylates, sebacinates, stearates, subacetates, succinates, sulfamineates, sulfates, tannates, tartrates, theoclates (8-chlorotheophylline), thiocyansides, triethiozides, undecanoates, undecylenates, and valersates.

[0056] Representative pharmaceutically acceptable base addition salts include, but are not limited to, aluminum, 2-amino-2-(hydroxymethyl)-1,3-propanediol (TRIS, tromethamine), arginine, benetamine (N-benzylphenethylamine), benzathine (N,N'-dibenzylethylenediamine), bis-(2-hydroxyethyl)amine, bismuth, calcium, chloroprocaine, choline, and cremisole (1-p-chlorobenzyl-2-pyrrolildine-1'-ylmethylbenzyl Examples include zuimidazole, cyclohexylamine, dibenzylethylenediamine, diethylamine, diethyltriamine, dimethylamine, dimethylethanolamine, dopamine, ethanolamine, ethylenediamine, L-histidine, iron, isoquinoline, lepidine, lithium, lysine, magnesium, meglumine (N-methylglucamine), piperazine, piperidine, potassium, procaine, quinine, quinoline, sodium, strontium, t-butylamine, and zinc.

[0057] A particularly preferred salt is the sodium salt. Another particularly preferred salt is the potassium salt.

[0058] In one embodiment, the composition of the present invention further comprises pharmaceutically acceptable additives. In the method of the present invention, preferred routes of administration are oral and intramuscular administration for subcutaneous or intramuscular delivery. Therefore, preferred pharmaceutical compositions include compositions suitable for oral administration (e.g., tablets) and compositions suitable for subcutaneous or intramuscular injection.

[0059] In another embodiment, the present invention discloses a method for preventing HIV infection in humans or reducing the risk of infection, comprising administering a pharmaceutical composition of the present invention. Pre-exposure prophylaxis (or PrEP) is when people at risk of HIV infection take medication daily to reduce their chances of HIV infection. PrEP has been shown to be effective in reducing the risk of infection.

[0060] The compounds and salts of the present invention are thought to have HIV capsids as their biological targets, and therefore their mechanism of action is to modify the function of HIV capsids in one or more ways.

[0061] The compounds and salts of the present invention can be used alone or in combination with other therapeutic agents. Accordingly, combination therapy according to the present invention comprises the administration of at least one compound or salt of the present invention and at least one other agent that may be useful in treating HIV infection. The compounds or salts of the present invention and other agents may be compounded and administered together in a single pharmaceutical composition, or they may be compounded and administered separately. If compounded and administered separately, they may be administered simultaneously or sequentially in any order. Other suitable drugs include, for example, abacavir, atazanavir, bictegravir, cabotegravir, darunavir, delavirdin, didanosine, dideoxyinosine, dolutegravir, doravirine, efavirenz, elvitegravir, emtricitabine, etavirine, fosanprenavir, fostemsavir, indinavir, slatravir, lamivudine, lopinavir, maraviroc, nelfinavir, nevirapine, raltegravir, rilpiverine, ritonavir, saquinavir, stabudine, tipranavir, tenofovir, tenofovir alafenamide, tenofovir disoproxil fumarate, zalcitabine, and zidovudine. Preferred drugs include, for example, islatravir, lamivudine, fostemsavir, and cabotegravir. Particularly preferred drugs include, for example, dolutegravir, bictegravir, lamivudine, fostemsavir, and cabotegravir. [Examples]

[0062] Preparation of bicyclo[3.1.0]hexane-3-ol

[0063] [ka]

[0064] Under an N2 atmosphere, at 0-5°C, a hexane solution of diethylzinc (1.0 M, 3091 mL, 3091 mmol) was added dropwise over 3 hours to a stirred DCM solution of cyclopent-3-enol (130 g, 1545 mmol). At 0°C, a DCM (300 mL) solution of diiodomethane (249 mL, 3091 mmol) was added dropwise over 1 hour. When the reaction mixture was heated to 27°C, the formation of a white precipitate was observed. The mixture was stirred for 16 hours. The progress of the reaction was monitored by TLC (SiO2, 20% siRNA / pet, Rf=0.3, UV-inactive, PMA-active). The reaction mixture was quenched by the careful addition of saturated NH4Cl aqueous solution (1.5 L). The mixture was filtered through a Celite pad. The aqueous layer was extracted with DCM (2 × 1 L). The combined organic layers were dried over anhydrous Na2SO4, filtered, and then concentrated under reduced pressure to obtain crude bicyclo[3.1.0]hexane-3-ol as a red liquid (180 g). 1 H NMR (400 MHz, CDCl3) δ = 4.41 - 4.35 (m, 1H), 2.18 - 2.05 (m, 2H), 1.73 (d, J = 13.9 Hz, 2H), 1.35 - 1.25 (m, 2H), 1.21 - 1.14 (m, 1H), 0.57 - 0.43 (m, 2H). GCMS: m / z = 98.1).

[0065] Preparation of bicyclo[3.1.0]hexane-3-one

[0066] [ka]

[0067] Under a N2 atmosphere at 0 °C, Dess-Martin periodinane (954 g, 225 mmol) was added portionwise to a stirred solution of bicyclo[3.1.0]hexan-3-ol (210 g, 2054 mmol) in DCM (5000 mL). The mixture was warmed to 27 °C and then stirred for 16 h. The progress of the reaction was monitored by TLC (SiO2, 20% acetone / Hex, Rf = 0.3, UV inactive, PMA active). The reaction mixture was filtered through a pad of Celite, and the filtrate was washed with aqueous NaOH solution (1 N, 8 × 1 L). The combined aqueous phases were extracted with DCM (5 × 1 L). The combined organic layers were dried over anhydrous Na2SO4, filtered, and then concentrated under reduced pressure (bath temperature: 20 °C) to give crude bicyclo[3.1.0]hexan-3-one as a brown liquid. The liquid was further purified by distillation at 70 °C to give bicyclo[3.1.0]hexan-3-one as a pale yellow viscous liquid (125 g, 62%). 1 1H NMR (400 MHz, CDCl3) δ = 2.61 - 2.54 (m, 2H), 2.17 - 2.12 (m, 2H), 1.54 - 1.46 (m, 2H), 0.92 - 0.86 (m, 1H), -0.01 - -0.08 (m, 1H); GCMS: M / Z = 96.1.

[0068] Preparation of 2-(2,2-difluoroacetyl)bicyclo[3.1.0]hexane-3-one

[0069]

Chemical formula

[0070] Under an N2 atmosphere, at -78°C, LDA (2.0 M THF solution, 0.701 L, 1402 mmol) was added to a stirred solution of bicyclo[3.1.0]hexane-3-one (125 g, 1274 mmol) in THF (1500 mL). The solution was stirred at -78°C for 1 hour. Difluoroethyl acetate (174 g, 1402 mmol) in THF (300 mL) was slowly added to the solution over 30 minutes while maintaining the temperature at -78°C. The reaction mixture was heated to 27°C and then stirred for 1 hour. The progress of the reaction was monitored by TLC (SiO2, 20% acetone / hexane, Rf=0.3, UV active). The reaction mixture was quenched by adding aqueous HCl (1 N, 2000 mL). The mixture was stirred for 30 minutes and then extracted with siRNA (3 × 1000 mL). The combined organic layers were washed with brine (1000 mL), dried over anhydrous sodium 2SO4, and filtered. The filtrate was concentrated under reduced pressure to obtain 2-(2,2-difluoroacetyl)bicyclo[3.1.0]hexane-3-one as a pale yellow viscous liquid (180 g, 71%). 1 H NMR (400 MHz, CDCl3) δ = 6.18 (t, J = 54.8 Hz, 1H), 2.70 - 2.62 (m, 1H), 2.35 (d, J = 19.4 Hz, 1H), 2.14 (br s, 1H), 1.26 - 1.21 (m, 1H), 1.04-1.03 (m, 1H), 0.22-0.21 (m, 1H), LCMS: M / Z = 173.17).

[0071] Preparation of ethyl 2-(3-(difluoromethyl)-3b,4,4a,5-tetrahydro-1H-cyclopropa[3,4]cyclopenta[1,2-c]pyrazole-1-yl) acetate

[0072] [ka]

[0073] Under an N2 atmosphere at 27°C, 2-(2,2-difluoroacetyl)bicyclo[3.1.0]hexane-3-one (180 g, 910 mmol) was stirred in ethanol (2 L), to which ethyl 2-hydrazinyl acetate hydrochloride (422 g, 2729 mmol) was added, followed by sulfuric acid (20 mL, 375 mmol). The mixture was stirred for 30 minutes, then heated to 100°C and stirred for 16 hours. The progress of the reaction was monitored by TLC (SiO2, 20% acetone / hexane, Rf=0.3, UV-active). The reaction mixture was concentrated under reduced pressure. The residue was dissolved in siRNA (2000 mL), washed with water (2 × 1 L) and brine (1 L), dried over anhydrous Na₂SO₄, filtered, and then concentrated under reduced pressure. The resulting residue was subjected to silica gel column chromatography (pet.:acetone 100:0 → 98:2) to obtain ethyl 2-(3-(difluoromethyl)-3b,4,4a,5-tetrahydro-1H-cyclopropa[3,4]cyclopenta[1,2-c]pyrazole-1-yl) acetate as an off-white solid (110 g, 46%). 1 H NMR (400 MHz, DMSO-d6) δ = 6.86 (t, J = 54.8 Hz, 1H), 4.93 (s, 2H), 4.14 (q, J = 7.2 Hz, 2H), 2.88 - 2.79 (m, 1H), 2.76 - 2.68 (m, 1H), 2.14 - 2.04 (m, 2H), 1.19 (t, J = 7.2 Hz, 3H), 1.10 - 1.03 (m, 1H), 0.14 (q, J = 4.3 Hz, 1H).

[0074] Preparation of ethyl 2-(3-(difluoromethyl)-5-oxo-3b,4,4a,5-tetrahydro-1H-cyclopropa[3,4]cyclopenta[1,2-c]pyrazole-1-yl) acetate

[0075] [ka]

[0076] At 0°C, pyridinium dichromate (794 g, 2110 mmol) was added in small increments to a stirred solution of ethyl 2-(3-(difluoromethyl)-3b,4,4a,5-tetrahydro-1H-cyclopropa[3,4]cyclopenta[1,2-c]pyrazole-1-yl) acetate (110 g, 422 mmol) and Celite (395 g) in cyclohexane (3.5 L). Under a nitrogen atmosphere, tert-butyl hydroperoxide (355 mL, 2130 mmol) was added dropwise to the mixture over 10 minutes. The reaction mixture was heated to 27°C and then stirred at that temperature for 48 hours. The progress of the reaction was monitored by TLC (SiO2, 30% acetone / pet, Rf=0.4, UV active). The reaction mixture was filtered, and the filter cake was extracted with ELISA (1000 mL). The filtrate was washed with saturated Na2S2O3 aqueous solution (2 × 500 mL), saturated FeSO4 aqueous solution (300 mL), and then brine (500 mL). The organic layer was dried over anhydrous Na2SO4, filtered, and concentrated under reduced pressure to obtain the crude title compound (150 g).

[0077] Preparation of ethyl 2-(3-(difluoromethyl)-4,4a-dihydrospiro[cyclopropa[3,4]cyclopenta[1,2-c]pyrazole-5,2'-[1,3]dithiolan]-1(3bH)-yl) acetate

[0078] [ka]

[0079] Under a nitrogen atmosphere at 27°C, ethyl 2-(3-(difluoromethyl)-5-oxo-3b,4,4a,5-tetrahydro-1H-cyclopropa[3,4]cyclopenta[1,2-c]pyrazole-1-yl) acetate (75 g, 269 mmol) was stirred in 1500 mL of DCM, to which ethane-1,2-dithiol (43.0 mL, 511 mmol) was added, followed by the addition of borotetraacetic acid trifluoride (72.6 mL, 511 mmol). The solution was stirred for 16 hours. The progress of the reaction was monitored by TLC (SiO2, 20% acetone / Pet, Rf=0.35, UV active). After completion, the reaction mixture was cooled to 0°C and quenched by adding saturated aqueous NaHCO3 solution (500 mL). The mixture was extracted with DCM (2 × 1000 mL). The combined organic matter was washed with brine (1000 mL), dried over anhydrous Na2SO4, filtered, and concentrated under reduced pressure to obtain a brown liquid. This substance was subjected to silica gel column chromatography (Pet.: siRNA 95:5 → 90:10) to obtain ethyl 2-(3-(difluoromethyl)-4,4a-dihydrospiro[cyclopropa[3,4]cyclopenta[1,2-c]pyrazole-5,2'-[1,3]dithiolan]-1(3bH)-yl) acetate as an off-white solid (80 g, 74%). 1 H-NMR (400 MHz, CDCl3) δ = 6.61 (t, J = 55.2 Hz, 1H), 5.00 - 4.85 (m, 2H), 4.29 - 4.19 (m, 2H), 3.55 - 3.46 (m, 4H), 2.63 - 2.53 (m, 1H), 2.49 - 2.38 (m, 1H), 1.30 - 1.24 (m, 4H), 0.65 - 0.60 (m, 1H). LCMS M+H = 346.9.

[0080] Preparation of ethyl 2-(3-(difluoromethyl)-5,5-difluoro-3b,4,4a,5-tetrahydro-1H-cyclopropa[3,4]cyclopenta[1,2-c]pyrazole-1-yl) acetate

[0081] [ka]

[0082] Under an N2 atmosphere, at -70°C, HF-pyridine (2.460 g, 24.83 mmol) was added to a stirred solution of 1,3-dibromo-5,5-dimethylimidazolidine-2,4-dione (26.3 g, 92 mmol) in DCM (20 mL). The solution was stirred for 30 minutes. To the solution, ethyl 2-(3-(difluoromethyl)-4,4a-dihydrospiro[cyclopropa[3,4]cyclopenta[1,2-c]pyrazole-5,2'-1,3]dithiolan]-1(3bH)-yl) acetate (10 g, 25 mmol) in DCM (20 mL) was added. The reaction mixture was heated to -40°C and then stirred at that temperature for 1 hour. The progress of the reaction was monitored by TLC (SiO2, 30% siRNA / PET, Rf=0.3, UV inactive). The reaction mixture was quenched by adding saturated NaHCO3 aqueous solution (200 mL). The mixture was heated to room temperature and then extracted with ethyl acetate (2 × 100 mL). The combined organic matter was washed with brine (50 mL), dried over anhydrous Na2SO4, filtered, and concentrated under reduced pressure to obtain a brown solid. This substance was subjected to silica gel column chromatography (Pet.: ethyl acetate 100: 0 → 75-25) to obtain ethyl 2-(3-(difluoromethyl)-5,5-difluoro-3b,4,4a,5-tetrahydro-1H-cyclopropa[3,4]cyclopenta[1,2-c]pyrazole-1-yl) acetate as a pale yellow solid (8.5 g, 91%). 1 H NMR (400 MHz, CDCl3) δ = 6.62 (t, J = 55.2 Hz, 1H), 4.82 (s, 2H), 4.30 - 4.18 (m, 2H), 2.51 - 2.37 (m, 2H), 1.42 - 1.35 (m, 1H), 1.31 - 1.23 (m, 3H), 1.14 - 1.08 (m, 1H). LCMS M+H = 293.07.

[0083] Preparation of 2-(3-(difluoromethyl)-5,5-difluoro-3b,4,4a,5-tetrahydro-1H-cyclopropa[3,4]cyclopenta[1,2-c]pyrazole-1-yl)acetic acid

[0084] [ka]

[0085] Under an N2 atmosphere at 0°C, ethyl 2-(3-(difluoromethyl)-5,5-difluoro-3b,4,4a,5-tetrahydro-1H-cyclopropa[3,4]cyclopenta[1,2-c]pyrazole-1-yl) acetate (15 g, 50 mmol) was mixed with THF (17 mL) and MeOH (66 mL) and then LiOH (1.788 g, 74.7 mmol) in aqueous solution (66 mL) was added. The reaction mixture was heated to 27°C and then stirred at that temperature for 3 hours. The progress of the reaction was monitored by TLC (SiO2, 5% MeOH / DCM, Rf=0.2, UV active). After completion, the reaction mixture was concentrated under reduced pressure, diluted with water (50 mL), and washed with ELISA (2 × 250 mL) to remove impurities. The aqueous layer was adjusted to pH 2-3 using an aqueous HCl solution (1M), and then extracted with HCl (3 × 1000 mL). The combined organic matter was dried over anhydrous Na₂SO₄, filtered, and concentrated under reduced pressure to obtain 2-(3-(difluoromethyl)-5,5-difluoro-3b,4,4a,5-tetrahydro-1H-cyclopropa[3,4]cyclopenta[1,2-c]pyrazole-1-yl)acetic acid as an off-white solid (14 g, 98%). LCMS M+H = 265.15.

[0086] Separation of the obtained 2-((3bS,4aR)-3-(difluoromethyl)-5,5-difluoro-3b,4,4a,5-tetrahydro-1H-cyclopropa[3,4]cyclopenta[1,2-c]pyrazole-1-yl)acetic acid and 2-((3bR,4aS)-3-(difluoromethyl)-5,5-difluoro-3b,4,4a,5-tetrahydro-1H-cyclopropa[3,4]cyclopenta[1,2-c]pyrazole-1-yl)acetic acid

[0087] [ka]

[0088] 5.5 g of 2-(3-(difluoromethyl)-5,5-difluoro-3b,4,4a,5-tetrahydro-1H-cyclopropa[3,4]cyclopenta[1,2-c]pyrazole-1-yl)acetic acid was dissolved in 20 mL of isopropanol. The solution was subjected to SFC chiral separation in small amounts as follows: Instrument = Thar 80; Column = Chiralpak IC 30 × 250 mm, 5 microns; Solvent A = Supercritical CO2; Solvent B = Isopropanol containing 0.5% isopropylamine (v / v); Eluent composition = 70% A:30% B; Flow rate = 65 g / min; Back pressure = 100 bar; Temperature = 30 °C; Injection volume = 2.5 mL; Detection = 220 nm. 2-((3bS,4aR)-3-(difluoromethyl)-5,5-difluoro-3b,4,4a,5-tetrahydro-1H-cyclopropa[3,4]cyclopenta[1,2-c]pyrazole-1-yl)acetic acid was recovered as a peak eluting at 7.5 to 14 minutes, and 2-((3bR,4aS)-3-(difluoromethyl)-5,5-difluoro-3b,4,4a,5-tetrahydro-1H-cyclopropa[3,4]cyclopenta[1,2-c]pyrazole-1-yl)acetic acid was recovered as a peak eluting at 2.7 to 5.8 minutes. For each enantiomer, the obtained solution was concentrated under reduced pressure, the resulting solid was dissolved in phenylethylamine, and then washed twice with 1M citric acid solution, followed by water, and then brine. The organic solution was dried with Na2SO4, filtered, and then concentrated under vacuum to obtain the separated enantiomer with a recovery rate of 80-90%.

[0089] Preparation of N-(7-amino-4-chloro-1-methyl-1H-indazole-3-yl)-N-(4-methoxybenzyl)methanesulfonamide

[0090] [ka]

[0091] Synthesis scheme

[0092] [ka]

[0093] Step 1: Preparation of 2,6-dichloro-3-nitrobenzaldehyde

[0094] [ka]

[0095] To a solution of sulfuric acid (H2SO4) (5.63 L, 4.5V) in a round-bottom flask at 0-5°C, 2,6-dichlorobenzaldehyde (1.25 kg, 7.10 mol, 1.0 equivalent) was gradually added at below 15°C. The reaction mixture was stirred at 0-5°C for 30 minutes. A freshly prepared solution of the nitration mixture [prepared at 0°C from concentrated H2SO4 (0.425 L, 0.34V) and 70% HNO3 (0.85 kg, 13.49 mol, 1.30 equivalent)] was added to the above reaction mixture at below 10°C [Note: This is a slightly exothermic reaction (3-6°C), therefore, addition at low temperatures is preferable]. The reaction mixture was stirred at 5-10°C for 2-3 hours. After the reaction was complete (monitored by TLC), the mixture was quenched at below 25°C with ice-cold water (18.75 L, 15V). Next, the reaction mixture was heated to room temperature and stirred for 2 hours. The solid was isolated by filtration and then washed with water (2.5 L, 2.0 V). Bulk residual water was removed from the solid by maintaining vacuum filtration for 60-90 minutes. The crudely moist solid was first dried in an air atmosphere. Then, the title product, 2,6-dichloro-3-nitrobenzaldehyde (1.44 kg, 92% yield), was dried in a hot air oven at 50-55°C for 10-12 hours (until the moisture content was less than 5.0%) to obtain a yellow solid. 1 H NMR (400 MHz, CDCl3): δ 10. 44 (s, 1H), 7.88 (d, J = 8.4 Hz, 1H), 7.56 (d, J = 8.8 Hz, 1H).

[0096] Step 2: Preparation of 2,6-dichloro-3-nitrobenzonitrile

[0097] [ka]

[0098] (Process-2a) 2,6-dichloro-3-nitrobenzaldehyde (1.17 kg, 5.31 mol, 1.0 equivalent) was added to a DMSO (5.9 L, 5.0 V) solution in a round-bottom flask at room temperature. After stirring at room temperature for 30 minutes, hydroxylamine hydrochloride (0.63 kg, 9.04 mol, 1.70 equivalents) was added, and the reaction mixture was stirred at room temperature for 3 hours. After the completion of the reaction (monitored by TLC), the reaction mixture was quenched by adding ice-cold water (18.0 L, 15.0 V) at a rate sufficient to maintain the temperature below 30°C (observation: solid formed upon water addition). The reaction mixture was stirred at room temperature for 60-90 minutes. The solid was isolated by filtration, washed with water (2.5 L, 2.0 V), and then washed with a mixture of acetone and hexane (6.0 L, 1:1 ratio). Bulk residual water was removed from the solid by maintaining vacuum filtration for 60-90 minutes. The moist solid was first air-dried, and then final-dried in a hot-air oven at 50-55°C for 10-12 hours (until the moisture content was less than 1.0%) to obtain the dried target product, 2,6-dichloro-3-nitrobenzaldehyde oxime (1.22 kg, 92% yield), as an off-white solid. The crude product (containing 10-20% 2,6-dichloro-3-nitrobenzonitrile) was used directly in the next step without further purification.

[0099] (Process-2b) At 0-5°C, triethylamine ("TEA", 1.02 kg, 10.09 mol, 2.1 equivalents) was added to a stirred solution of crude oxime (the above preparation, 1.13 kg, 4.80 mol, 1.0 equivalent) in DCM (9.04 L, 8.0 V). After stirring for 5 minutes, methanesulfonyl chloride (0.60 kg, 5.29 mol, 1.1 equivalents) was slowly added at 15°C (observation: exothermic reaction was observed during addition). The reaction mixture was then stirred at room temperature for 30-45 minutes. After the reaction was complete (the progress of the reaction was monitored by TLC; mobile phase: hexane solution of 20% ethyl acetate), the reaction mixture was diluted with water (6.78 L, 6.0 V), the organic layer was separated, and the aqueous layer was extracted with DCM (3.4 L, 3.0 V). The combined organic layers were washed with brine (5.65 L, 5.0 V), dried over Na2SO4, and concentrated under vacuum. The obtained crude solid was triturated with hexane (4.50 L, 4.0 V) at room temperature. The wet material was dried in a hot air oven at 50-55°C for 5-6 hours to obtain the dried product, 2,6-dichloro-3-nitrobenzonitrile (0.95 kg, 91% yield), as a yellow solid. 1 H NMR (400 MHz, CDCl3): δ 8.07 (d, J = 8.8 Hz, 1H), 7.63 (d, J = 8.8 Hz, 1H).

[0100] Step 3: Preparation of 4-chloro-7-nitro-1H-indazole-3-amine

[0101] [ka]

[0102] At 15-20°C, hydrazine hydrate (519.0 g, 10.36 mol, 3.0 equivalents) was slowly added to a stirred solution of 2,6-dichloro-3-nitrobenzonitrile (750.0 g, 3.45 mol, 1.0 equivalent) in ethanol (7.5 L, 10.0 V), while maintaining the reaction mixture below 25°C (Observation: slight exothermic reaction occurred upon addition, and solid formation began upon addition). The temperature of the reaction mixture was slowly raised to room temperature, and the mixture was then stirred for 3 hours (Observation: the amount of solid increased during this time). After the reaction was complete (monitored by TLC), the mixture was diluted with water (7.5 L, 10.0 V) and stirred at room temperature for another hour. The solid was isolated by filtration and then washed with water (2.25 L, 3.0 V). The wet solid was washed with a 1:1 mixture of acetone (1.875 L, 2.5V) and hexane (1.875 L, 2.5V). Bulk residual water was removed from the solid by maintaining vacuum filtration for 60–90 minutes. Finally, the wet solid was dried in a hot air oven at 50°C for 7–8 hours (until the moisture content was less than 1.5%) to obtain the dried product, 4-chloro-7-nitro-1H-indazole-3-amine (549.0 g, 75% yield), as a brick-red solid. 1 H NMR (400 MHz, CDCl3): δ 10.36 (bs, 1H), 8.20 (d, J = 8.4 Hz, 1H), 7.07 (d, J = 8.40 Hz, 1H), 4.73 (bs, 2H).

[0103] Step 4: Preparation of 4-chloro-1-methyl-7-nitro-1H-indazole-3-amine

[0104] [ka]

[0105] At 5-10°C, cesium carbonate (Cs2CO3) (1.91 kg, 5.88 mol, 2.5 equivalents) was slowly added to a stirred solution of 4-chloro-7-nitro-1H-indazole-3-amine (500 g, 0.42 mol, 1.0 equivalent) in DMF (5.0 L, 10.0 V), while maintaining the reaction mixture below 10°C. After stirring for 5-10 minutes, dimethyl sulfate (326.3 g, 2.59 mol, 1.1 equivalents) was added while maintaining the reaction mixture below 10°C (Note: For better regioselectivity, slow addition is preferable). The reaction temperature was then slowly raised to room temperature, and stirring was continued at the same temperature for a further 2 hours. After the completion of the reaction (monitored by TLC), the reaction mixture was quenched by adding ice-cold water (15.0 L, 30.0 V), and the resulting mixture was stirred at room temperature for 6-8 hours. The solid was isolated by filtration and then washed with water (1.5 L, 3.0 V). The wet solid was washed with IPA (1.5 L, 3.0 V) and then with hexane (1.0 L, 2.0 V). Bulk residual water was removed from the solid by maintaining vacuum filtration for 60-90 minutes. The wet solid was dried in a 50°C hot air oven for 7-8 hours (until the moisture content was less than 1.0%). The isolated substance, 4-chloro-1-methyl-7-nitro-1H-indazole-3-amine (319.0 g, 60% yield), was used in the next step without further purification. 1 H NMR (400 MHz, CDCl3): δ 7.97 (d, J = 8.32 Hz, 1H), 6.97 (d, J = 8.24 Hz, 1H), 4.63 (bs, 2H), 3.96 (s, 3H).

[0106] Step 5: Preparation of N-(4-chloro-1-methyl-7-nitro-1H-indazole-3-yl)methanesulfonamide

[0107] [ka]

[0108] (Step 5a) At 0-5°C, triethylamine (TEA) (837.0 g, 8.27 mol, 3.0 equivalents) was added to a solution of 4-chloro-1-methyl-7-nitro-1H-indazole-3-amine (625.0 g, 2.76 mol, 1.0 equivalent) in DCM (6.25 L, 10.0 V), followed by 4-dimethylaminopyridine (DMAP) (20.60 g, 0.165 mol, 0.06 equivalents). The reaction mixture was stirred for 5-10 minutes, and then methanesulfonyl chloride (MsCl) (790.0 g, 6.89 mol, 2.5 equivalents) was slowly added while maintaining the reaction mixture below 10°C. The reaction mixture was warmed to room temperature and then stirred for 1.5-2.0 hours. After the reaction was complete (monitored by TLC), the mixture was diluted with water (6.25 L, 10.0 V) and then stirred at room temperature for 15 minutes. The organic layer was separated, and the aqueous layer was extracted with DCM (6.25 L, 10.0 V). The combined organic layers were washed with brine (1.25 L, 2.0 V), dried over Na2SO4, and concentrated to obtain a crude solid. The solid was triturated with hexane (1.25 L, 2.0 V) at room temperature to obtain the intermediate N-(4-chloro-1-methyl-7-nitro-1H-indazole-3-yl)-N-(methylsulfonyl)methanesulfonamide, which was used directly in the next step.

[0109] (ii) At room temperature, 4.38 L of 5% NaOH aqueous solution (7.0 V) was slowly added to a stirred solution of N-(4-chloro-1-methyl-7-nitro-1H-indazole-3-yl)-N-(methylsulfonyl)methanesulfonamide (prepared above) in ethanol (20.0 V) [Note: It is preferable to add slowly via a dropping funnel]. The reaction mixture was stirred at the same temperature for 3 hours. After the reaction was complete (monitored by TLC) [Sample preparation for TLC analysis: Acidify approximately 1.0 mL of the sample with 2.0 N HCl aqueous solution to pH 2-3, extract with ethyl acetate, and analyze the organic layer by TLC]. The reaction mixture was cooled to 0-5°C. While maintaining the reaction temperature below 10°C, the pH was adjusted to 2-3 by adding 3.13 L of 2.0 N HCl aqueous solution (5.0 V) [Note: Precipitation occurred upon addition of HCl and increased with stirring]. The reaction mixture was heated to room temperature and then stirred for 1.5–2.0 hours. The resulting solid was isolated by filtration, washed with water (1.25 L, 2.0 V), and then washed with hexane (1.25 L, 2.0 V). Bulk residual water was removed from the solid by maintaining vacuum filtration for 60–90 minutes. The wet material was dried in a 50°C hot air oven for 6–7 hours (until the moisture content was less than 1.0%) to obtain the dried product N-(4-chloro-1-methyl-7-nitro-1H-indazole-3-yl)methanesulfonamide (640.0 g, 76%) as a yellow solid. 1 H NMR (400 MHz, CDCl3): δ 8.05 (d, J = 8.32 Hz, 1H), 7.32 (bs, 1H), 7.17 (d, J = 8.28 Hz, 1H), 4.15 (s, 3H), 3.45 (s, 3H).

[0110] Step 6: Preparation of N-(4-chloro-1-methyl-7-nitro-1H-indazole-3-yl)-N-(4-methoxybenzyl)methanesulfonamide

[0111] [ka]

[0112] At room temperature, potassium carbonate (374.7 g, 2.70 mol, 1.3 equivalents) was added to a solution of N-(4-chloro-1-methyl-7-nitro-1H-indazole-3-yl)methanesulfonamide (635.0 g, 2.08 mol, 1.0 equivalent) and 1-(chloromethyl)-4-methoxybenzene (359.0 g, 2.30 mol, 1.1 equivalents) in DMF (6.35 L, 10.0 V). The reaction mixture was heated to 80-90°C and maintained at that temperature for 3 hours. After the reaction was complete (monitored by TLC), the mixture was poured into ice-cold water (19.05 L, 30.0 V) [Note: It is preferable to quench slowly with vigorous stirring to avoid aggregation as the product precipitates]. The obtained solid was isolated by filtration, washed with water (1.90 L, 3.0 V), and then washed with hexane (1.27 L, 2.0 V). Bulk residual water was removed from the solid by maintaining vacuum filtration for 60-90 minutes. The isolated solid was dissolved in ethyl acetate (12.7 L, 20.0 V), and charcoal (63.5 g) was added. The mixture was heated to 60-70°C and then stirred at that temperature for 30-45 minutes. While still hot (40-50°C), the mixture was filtered through a Celite pad, and then the Celite pad was extracted with ethyl acetate (3.17 L, 5.0 V). The combined filtrate was concentrated to dryness under reduced pressure at less than 50°C. Ethyl acetate (0.635 L, 1.0 V) was added to the solid at room temperature. The resulting solid suspension was stirred for 30 minutes. The solid was isolated by filtration and then washed with hexane (1.27 L, 2.0 V). By maintaining vacuum filtration for 45-60 minutes, residual water was removed from the solid to obtain the product N-(4-chloro-1-methyl-7-nitro-1H-indazole-3-yl)-N-(4-methoxybenzyl)methanesulfonamide (705.0 g, 80% yield) as a yellow solid. 1 H NMR (400 MHz, CDCl3): δ 7.99 (d, J = 8.24 Hz, 1H), 7.27 (d, J = 8.68 Hz, 2H), 7.19 (d, J = 8.24 Hz, 1H), 6.80 (d, J = 8.44 Hz, 2H), 4.95-4.76 (m, 2H), 4.17 (s, 3H), 3.76 (s, 3H), 3.01 (s, 3H).

[0113] Step 7: Preparation of N-(7-amino-4-chloro-1-methyl-1H-indazole-3-yl)-N-(4-methoxybenzyl)methanesulfonamide

[0114] [ka]

[0115] At room temperature, ammonium chloride (NH4Cl) (449.0 g, 8.23 ​​mol, 10.0 equivalents) was added to a stirred suspension of zinc powder (540.0 g, 8.23 ​​mol, 10.0 equivalents) in a mixture of THF (3.50 L, 10.0 V) and water (7.0 L, 20.0 V). To the mixture, a solution of N-(4-chloro-1-methyl-7-nitro-1H-indazole-3-yl)-N-(4-methoxybenzyl)methanesulfonamide (350 g, 0.823 mol, 1.0 equivalent) in THF (7.0 L, 20.0 V) was added. The reaction mixture was stirred at room temperature for 3-4 hours. After the reaction was complete (monitored by in-process TLC / HPLC), the mixture was diluted with ethyl acetate (3.5 L, 10.0 V) and water (1.12 L, 2.5 V). The mixture was stirred for 15 minutes. The reaction mixture was filtered through a Celite bed pad washed with ethyl acetate (1.75 L, 5.0 V). The biphasic filtrate was collected and the phases were separated. The aqueous layer was extracted with ethyl acetate (3.50 L, 10.0 V). The combined organic layers were washed with brine (3.50 L, 10 V), dried over Na2SO4, and then concentrated under vacuum to obtain a crude solid. MTBE (3.25 L, 10 V) was added to the crude product, and the suspension was stirred at room temperature for 30 minutes. The solid was isolated by filtration. Bulk residual water was removed from the solid by maintaining vacuum filtration for 30–45 minutes. The wet product was dried in a hot air oven (50°C) for 2 hours to obtain the title product N-(7-amino-4-chloro-1-methyl-1H-indazole-3-yl)-N-(4-methoxybenzyl)methanesulfonamide (276.0 g, 85% yield) as an off-white solid. 1H NMR (400 MHz, CDCl3): δ 7.29-7.26 (m, 2H), 6.86-6.79 (m, 2H), 6.42 (d, J = 7.80 Hz, 1H), 4.99-4.70 (m, 2H), 4.25 (s, 3H), 3.77 (s, 5H), 2.98 (s, 3H).

[0116] Preparation of 2-amino-4-(6-(trifluoromethyl)pyridine-2-yl)benzoic acid

[0117] [ka]

[0118] Synthesis scheme

[0119] [ka]

[0120] Step 1: Preparation of methyl 4-bromo-2-nitrobenzoate

[0121] 500 mL of sulfuric acid (9381 mmol) was added at 0°C to a stirred solution of 4-bromo-2-nitrobenzoic acid (500 g, 2032 mmol) in methanol (2000 mL). The solution was stirred at 70°C for 4 hours. The progress of the reaction was monitored by TLC (SiO2, 30% siRNA / Pet. Rf=0.3). After the reaction was complete, the reaction mixture was cooled to room temperature and then concentrated under reduced pressure to remove methanol. The resulting residue was poured into water (1000 mL) and the pH was adjusted to pH 9 by adding anhydrous sodium carbonate. The mixture was extracted with ethyl acetate (2 × 1000 mL). The combined organic matter was washed with water (500 mL) and then with brine solution (500 mL). The organic layer was dried over anhydrous Na2SO4, filtered, and then concentrated under reduced pressure to obtain methyl 4-bromo-2-nitrobenzoate (520 g, 94%) as an off-white solid. ¹H-NMR (400 MHz, CDCl₃) δ = 8.04–7.99 (m, ¹H), 7.85–7.78 (m, ¹H), 7.66 (d, J = 8.2 Hz, ¹H), 3.92 (s, ³H). LCMS Purity = 95.2%. The product was used directly in the next step without further purification.

[0122] Step 2: Preparation of methyl 2-amino-4-bromobenzoate To a stirred solution of zinc powder (704 g, 10.8 mol) in water (2000 mL), a solution of methyl 4-bromo-2-nitrobenzoate (400 g, 1538 mmol) in tetrahydrofuran (THF) (4000 mL) was slowly added under a nitrogen atmosphere at 0°C, followed by the slow addition of acetic acid (1057 mL, 18.5 mol). The reaction mixture was stirred at 27°C for 4 hours. The progress of the reaction was monitored by TLC (SiO2, 20% ethyl acetate / Pet. Rf=0.4). Upon completion, the reaction mixture was filtered through a Celite pad, and the Celite pad was extracted with ethyl acetate (2000 mL). The combined filtrate was concentrated under reduced pressure. The resulting residue was diluted with water (3000 mL) and extracted with ethyl acetate (2 × 4000 mL). The combined organic matter was washed with saturated Na2CO3 solution (2 × 3000 mL), followed by brine (2 × 2000 mL). The organic layer was dried over anhydrous Na2SO4, filtered, and then concentrated under reduced pressure to obtain methyl 2-amino-4-bromobenzoate (350 g, 94%) as an off-white solid. ¹H-NMR (400 MHz, CDCl3) δ = 7.76–7.65 (m, 1H), 6.84 (d, J = 1.9 Hz, 1H), 6.78–6.72 (m, 1H), 5.91–5.63 (m, 2H), 3.86 (s, 3H). LCMS Purity = 95.0%. The product was used directly in the next step without further purification.

[0123] Step 3: Preparation of methyl 2-amino-4-(6-(trifluoromethyl)pyridine-2-yl)benzoate

[0124] [ka]

[0125] To a stirred solution of methyl 2-amino-4-bromobenzoate (350 g, 1521 mmol) in 1,4-dioxane (7000 mL), bis(pinacol)diborane (522 g, 2054 mmol) and potassium acetate (597 g, 6085 mmol) were added. The reaction mixture was degassed by bubbling N2 gas through it for 10 minutes. PdCl2 (dppf) (78 g, 106 mmol) was added to the reaction mixture. The mixture was stirred at 90°C for 4 hours. The progress of the reaction was monitored by TLC. Once the reaction was complete, the mixture was cooled to room temperature. 2-chloro-6-(trifluoromethyl)pyridine (359 g, 1978 mmol), tribasic potassium phosphate (1130 g, 5325 mmol) and water (1190 mL) were added to the mixture. The mixture was degassed by bubbling nitrogen gas for 10 minutes. PdCl2 (dppf) (78 g, 106 mmol) was added to the mixture. The reaction mixture was stirred at 60°C for 16 hours. The progress of the reaction was monitored by TLC (SiO2, 20% siRNA / PET, Rf=0.4). Upon completion, the reaction mixture was filtered through Celite, and the Celite pad was extracted with ethyl acetate (2000 mL). The combined filtrate was concentrated under reduced pressure to obtain the crude product (550 g) as a brown liquid. This substance was purified by silica gel chromatography eluting with 5-30% siRNA / PET. The fraction containing the desired product was pooled and concentrated under reduced pressure. The isolated substance was washed with n-pentane (2200 mL), the solid was recovered by filtration, and then dried under vacuum to obtain methyl 2-amino-4-(6-(trifluoromethyl)pyridine-2-yl)benzoate (380 g, 83%) as an off-white solid. 1 H NMR (CHLOROFORM-d) δ: 7.99 (d, J=8.3 Hz, 1H), 7.92-7.98 (m, 2H), 7.67 (dd, J=6.9, 1.5 Hz, 1H), 7.49 (d, J=1.5 Hz, 1H), 7.27 (dd, J=8.3, 1.8 Hz, 1H), 5.90 (br s, 2H), 3.93 (s, 3H). LCMS Purity = 98.25%.

[0126] Step 4: Preparation of 2-amino-4-(6-(trifluoromethyl)pyridine-2-yl)benzoic acid Under a nitrogen atmosphere at 0°C, lithium hydroxide monohydrate (369 g, 8776 mmol) was added to a stirred solution of methyl 2-amino-4-(6-(trifluoromethyl)pyridine-2-yl)benzoate (650 g, 2194 mmol) in tetrahydrofuran (THF) (5000 mL) and water (2167 mL). The reaction mixture was stirred at 70°C for 16 hours. The progress of the reaction was monitored by TLC (SiO2, 50% Â / Pet. Rf=0.4). Upon completion, the reaction mixture was concentrated under reduced pressure, and the resulting residue was dissolved in water (5000 mL) and acidified to pH 4 with the addition of 3N HCl (3000 mL). The resulting precipitate was filtered, washed with water (4000 mL), then with n-hexane (5000 mL), and then dried to obtain 2-amino-4-(6-(trifluoromethyl)pyridine-2-yl)benzoic acid (581 g, 93%) as an off-white solid. ¹H-NMR (400 MHz, DMSO-d6) δ = 8.18 (d, J = 4.0 Hz, 2H), 7.92-7.88 (m, 1H), 7.84 (d, J = 8.3 Hz, 1H), 7.54 (d, J = 1.7 Hz, 1H), 7.22 (d, J = 8.4, 1.8 Hz, 1H). LCMS Purity = 99.62%.

[0127] Example 1: Preparation of N-((S)-1-((3P)-3-(4-chloro-1-methyl-3-(methylsulfonamide)-1H-indazole-7-yl)-4-oxo-7-(6-(trifluoromethyl)pyridine-2-yl)-3,4-dihydroquinazoline-2-yl)-2-(3,5-difluorophenyl)ethyl)-2-((3bS,4aR)-3-(difluoromethyl)-5,5-difluoro-3b,4,4a,5-tetrahydro-1H-cyclopropa[3,4]cyclopenta[1,2-c]pyrazole-1-yl)acetamide

[0128] [ka]

[0129] Synthesis scheme

[0130] [ka]

[0131] Step 1: Preparation of tert-butyl(S)-(1-(3-(4-chloro-3-(N-(4-methoxybenzyl)methylsulfonamide)-1-methyl-1H-indazole-7-yl)-4-oxo-7-(6-(trifluoromethyl)pyridine-2-yl)-3,4-dihydroquinazoline-2-yl)-2-(3,5-difluorophenyl)ethyl)carbamate Under a nitrogen atmosphere at 27°C, pyridine (0.059 L, 724 mmol) was added to a stirred solution of (S)-2-((tert-butoxycarbonyl)amino)-3-(3,5-difluorophenyl)propanoic acid (91 g, 301 mmol) and 2-amino-4-(6-(trifluoromethyl)pyridine-2-yl)benzoic acid (94 g, 332 mmol) in acetonitrile (3.8 L) at 27°C. The resulting mixture was cooled to -9°C for 10 minutes, and then 2,4,6-tripropyl-1,3,5,2,4,6-trioxatriphosfinan 2,4,6-trioxide ("T3P", 50 wt% Â solution, 0.888 L, 1507 mmol) was added dropwise over 10 minutes. The solution was stirred under a N2 atmosphere at -9°C for 2.1 hours. N-(7-amino-4-chloro-1-methyl-1H-indazole-3-yl)-N-(4-methoxybenzyl)methanesulfonamide (120 g, 301 mmol) was added to a solution at -9°C. The solution was then heated to -5°C and maintained at that temperature with stirring for 1 hour. The reaction mixture was then slowly heated to 27°C and stirred at that temperature for 16 hours. The progress of the reaction was monitored by TLC (SiO2, 50% siRNA / Pet. Rf=0.5). Upon completion, the reaction mixture was concentrated under reduced pressure, the resulting residue was dissolved in siRNA (5000 mL), then washed with 1N NaOH solution (2000 mL), and then brine (1000 mL). The organic layer was dried over Na2SO4, filtered, and then concentrated under reduced pressure to obtain the crude product, which was purified by silica gel chromatography eluted with 30-35% siRNA / Pet. The fraction containing the desired product was pooled and concentrated under reduced pressure to obtain tert-butyl(S)-(1-(3-(4-chloro-3-(N-(4-methoxybenzyl)methylsulfonamide)-1-methyl-1H-indazole-7-yl)-4-oxo-7-(6-(trifluoromethyl)pyridine-2-yl)-3,4-dihydroquinazoline-2-yl)-2-(3,5-difluorophenyl)ethyl)carbamate (252 g, 88%, off-white solid) as a mixture of homochiral atropisomers (diastereomers).

[0132] Step 2: (S)-N-((6P)-7-(2-(1-amino-2-(3,5-difluorophenyl)ethyl)-4-oxo-7-(6-(trifluoromethyl)pyridine-2-yl)quinazoline-3(4H)-yl)-4-chloro-1-methyl-1H-indazole-3-yl)methanesulfonamide At 27°C, trifluoromethanesulfonic acid (70.4 mL, 793 mmol) was added to a stirred solution of tert-butyl(S)-(1-(3-(4-chloro-3-(N-(4-methoxybenzyl)methylsulfonamide)-1-methyl-1H-indazole-7-yl)-4-oxo-7-(6-(trifluoromethyl)pyridine-2-yl)-3,4-dihydroquinazolin-2-yl)-2-(3,5-difluorophenyl)ethyl) carbamate (97% purity, 252 g, 264 mmol) in TFA (815 mL, 10.6 mol). The solution was stirred under a nitrogen atmosphere for 2 hours. The progress of the reaction was monitored by TLC (SiO2, 50% Â / Pet. Rf=0.2). Upon completion, volatile substances were removed under a gentle stream of nitrogen gas. The residue was dissolved in ELISA (5000 mL), then washed with 1N NaOH solution (2000 mL), and then brine (1500 mL). The organic layer was dried over Na2SO4, filtered, and then concentrated under reduced pressure to obtain the crude product, which was purified by silica gel chromatography eluting with 5-15% MeOH DCM solution. The fraction containing the desired product was pooled and concentrated under reduced pressure to obtain (S)-N-(7-(2-(1-amino-2-(3,5-difluorophenyl)ethyl)-4-oxo-7-(6-(trifluoromethyl)pyridine-2-yl)quinazoline-3(4H)-yl)-4-chloro-1-methyl-1H-indazole-3-yl)methanesulfonamide (180 g, 95%, off-white solid) as a mixture of homochiral atropisomers (diastereomers). This substance was dissolved in methanol:acetonitrile (40:60, 3000 mL) and then purified by preparative SFC using the following method (column = (R,R)Welk-01, 30 × 250 mm, 5 μm, eluent = CO2:methanol (1:1); flow rate = 90.0 g / min; back pressure = 120.0 bar; detection = 254 nm (UV); stack time = 8.8 min; input per injection = 700 mg). Two peaks were generated by SFC separation and recovered separately.The major peak (the second peak to elute) was concentrated under reduced pressure to obtain (S)-N-((6P)-7-(2-(1-amino-2-(3,5-difluorophenyl)ethyl)-4-oxo-7-(6-(trifluoromethyl)pyridine-2-yl)quinazoline-3(4H)-yl)-4-chloro-1-methyl-1H-indazole-3-yl)methanesulfonamide (100 g, 54%) as an off-white solid. The product is a single stereoisomer. 1H-NMR (400 MHz, DMSO-d6) δ = 8.64-8.55 (m, 2H), 8.44-8.25 (m, 3H), 8.01 (d, J = 7.7 Hz, 1H), 7.42-7.31 (m, 2H), 7.07-6.95 (m, 1H), 6.76 (dd, J = 2.0, 8.5 Hz, 2H), 3.70 (s, 3H), 3.59 (dd, J = 4.8, 8.2 Hz, 1H), 3.35 (br d, J = 4.8 Hz, 1H), 3.17 (d, J = 5.1 Hz, 3H), 2.92-2.83 (m, 1H).LCMS Purity = 99%.

[0133] Step 3: Preparation of N-((S)-1-((3P)-3-(4-chloro-1-methyl-3-(methylsulfonamide)-1H-indazole-7-yl)-4-oxo-7-(6-(trifluoromethyl)pyridine-2-yl)-3,4-dihydroquinazoline-2-yl)-2-(3,5-difluorophenyl)ethyl)-2-((3bS,4aR)-3-(difluoromethyl)-5,5-difluoro-3b,4,4a,5-tetrahydro-1H-cyclopropa[3,4]cyclopenta[1,2-c]pyrazole-1-yl)acetamide At 27°C, (S)-N-((6P)-7-(2-(1-amino-2-(3,5-difluorophenyl)ethyl)-4-oxo-7-(6-(trifluoromethyl)pyridine-2-yl)quinazolin-3(4H)-yl)-4-chloro-1-methyl-1H-indazole-3-yl)methanesulfonamide (45g, 63.9 mmol) is mixed with DMF (450mL) and 2-((3bS,4aR)-3-(difluoromethyl)-5,5-difluoro-3b,4 4a,5-tetrahydro-1H-cyclopropa[3,4]cyclopenta[1,2-c]pyrazole-1-yl)acetic acid (20.26 g, 77 mmol), followed by N-(3-dimethylaminopropyl)-N'-ethylcarbodiimide hydrochloride ("EDC-HCl", 14.70 g, 77 mmol), 1-hydroxybenzotriazole hydrate ("HOBt hydrate", 11.75 g, 77 mmol), and N-methylmorpholine (28.1 mL, 256 mmol). The reaction mixture was stirred at 27°C for 24 hours. The progress of the reaction was monitored by TLC (SiO2, 50% siRNA / Pet. Rf=0.5). Upon completion, the reaction mixture was diluted with ice water (1.5 L), the resulting precipitate was collected by filtration, and then dried under vacuum to obtain the crude product (59 g) as an off-white solid. This crude product was blended with another batch (61 g) of crude product produced by repeating the procedure on the same scale. The combined 120 g of crude product was purified by silica gel chromatography eluting with 20-40% siRNA / PET. The fraction containing the desired product was pooled and concentrated under reduced pressure to obtain the purified product.The compound was ground using a mortar and pestle, and then trace amounts of HCl residue were removed by holding the fine solid in a 50°C oven for approximately 2 hours. The grinding and heating process was repeated four more times until the ethyl alcohol content was reduced to less than 4000 ppm to obtain N-((S)-1-((3P)-3-(4-chloro-1-methyl-3-(methylsulfonamide)-1H-indazole-7-yl)-4-oxo-7-(6-(trifluoromethyl)pyridine-2-yl)-3,4-dihydroquinazoline-2-yl)-2-(3,5-difluorophenyl)ethyl)-2-((3bS,4aR)-3-(difluoromethyl)-5,5-difluoro-3b,4,4a,5-tetrahydro-1H-cyclopropa[3,4]cyclopenta[1,2-c]pyrazole-1-yl)acetamide (85.7 g, 79%) as an off-white solid. 1H NMR (acetone-d6) δ: 8.60 (t, J=1.0 Hz, 1H), 8.58 (s, 1H), 8.48 (d, J=8.0 Hz, 1H), 8.39 (d, J=0.9 Hz, 2H), 8.32 (t, J=7.9 Hz, 1H), 8.16 (d, J=8.6 Hz, 1H), 7.95-7.99 (m, 1H), 7.49 (d, J=7.7 Hz, 1H), 7.37 (d, J=8.0 Hz, 1H), 6.86 (tt, J=9.3, 2.3 Hz, 1H), 6.70-6.76 (m, 2H), 6.77 (t, J=54.7 Hz, 1H), 4.93 (td, J=9.0, 4.6 Hz, 1H), 4.63-4.74 (m, 2H), 3.69 (s, 3H), 3.55 (dd, J=14.2, 4.6 Hz, 1H), 3.28 (s, 3H), 3.15 (dd, J=14.2, 9.4 Hz, 1H), 2.42-2.51 (m, 2H), 1.37-1.43 (m, 1H), 0.95-1.00 (m, 1H).LCMS analysis method: Column: Acquity BEH C18, 2.1 × 50 mm, 1.7 μm particles; Solvent A = 0.1% formic acid aqueous solution; Solvent B = 0.1% formic acid in acetonitrile solution; Flow rate = 0.6 mL / min; Gradient {Time point (min) / %B (%) at time point} = 0 / 3, 0.4 / 3, 7.5 / 98, 9.5 / 98, 9.6 / 3, 10 / 3; Column temperature = 35°C. LCMS results: Retention time = 5.57 min; Observed ions = 949.98 (M + H); LCMS purity = 99.4%.

[0134] Naming of Example 1: The compound of Example 1 prepared above is a homochiral substance containing axial chirality. Axial chirality can be described using the O / M nomenclature detailed in the IUPAC Gold Book (doi:10.1351 / goldbook.A00547). However, at present, the number of software tools capable of generating chemical names including P / M nomenclature is limited, and the options for converting chemical names using this nomenclature into molecular structural representations are even more limited. Therefore, for clarity and convenience, some of the names of Example 1 are shown below.

[0135] The names of the examples generated by ChemDraw Professional 16 (without P / M nomenclature) are as follows: N-((S)-1-(3-(4-chloro-1-methyl-3-(methylsulfonamide)-1H-indazole-7-yl)-4-oxo-7-(6-(trifluoromethyl)pyridine-2-yl)-3,4-dihydroquinazoline-2-yl)-2-(3,5-difluorophenyl)ethyl)-2-((3bS,4aR)-3-(difluoromethyl)-5,5-difluoro-3b,4,4a,5-tetrahydro-1H-cyclopropa[3,4]cyclopenta[1,2-c]pyrazole-1-yl)acetamide

[0136] The chemical names of Example 1, generated by JChem for Excel (including P / M nomenclature), are as follows: N-[(1S)-1-[3-(4-chloro-3-methanesulfonamide-1-methyl-1H-indazole-7-yl)-4-oxo-7-[6-(trifluoromethyl)pyridine-2-yl]-3,4-dihydroquinazoline-2-yl]-2-(3,5-difluorophenyl)ethyl]-2-[(2S,4R)-9-(difluoromethyl)-5,5-difluoro-7,8-diazatricyclo[4.3.0.0 2 , 4 ]nona-1(6),8-dien-7-yl]acetamide

[0137] The chemical names of Example 1, generated by ChemDraw Professional 16 with manual addition of P / M nomenclature, are as follows: N-((S)-1-((3P)-3-(4-chloro-1-methyl-3-(methylsulfonamide)-1H-indazole-7-yl)-4-oxo-7-(6-(trifluoromethyl)pyridine-2-yl)-3,4-dihydroquinazoline-2-yl)-2-(3,5-difluorophenyl)ethyl)-2-((3bS,4aR)-3-(difluoromethyl)-5,5-difluoro-3b,4,4a,5-tetrahydro-1H-cyclopropa[3,4]cyclopenta[1,2-c]pyrazole-1-yl)acetamide

[0138] Preparation of 2-amino-4-(4-(difluoromethyl)pyrimidine-2-yl)benzoic acid [ka]

[0139] Synthesis scheme

[0140] [ka]

[0141] Step 1: Preparation of methyl 2-amino-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)benzoate Under argon, a mixture of methyl 2-amino-4-bromobenzoate (10 g, 43.5 mmol), 4,4,4',4',5,5,5',5'-octamethyl-2,2'-bi(1,3,2-dioxaborolane) (11.04 g, 43.5 mmol), PdCl2 (dppf) (1.590 g, 2.173 mmol), and potassium acetate (12.80 g, 130 mmol) in 1,4-dioxane (100 mL) was heated at 97°C for 2 hours. The mixture was cooled to room temperature and then diluted with DCM. The organic layer was washed with water and then brine, dried over MgSO4, filtered, and concentrated under reduced pressure to obtain the crude product. The crude product was purified by silica gel chromatography (330 g column, 5-30% siRNA:Hex) to obtain methyl 2-amino-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)benzoate (9.2 g, 76%) as a yellow solid. 1 H NMR (500 MHz, CDCl3) δ ppm 7.86 (d, J=8.05 Hz, 1 H), 7.14 (s, 1 H), 7.06 (dd, J=7.90, 1.04 Hz, 1 H), 5.67 (br s, 2 H), 3.89 (s, 3 H), 1.37 (s, 12 H).

[0142] Step 2: Preparation of methyl 2-amino-4-(4-(difluoromethyl)pyrimidine-2-yl)benzoate In a round-bottom flask, methyl 2-amino-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)benzoate (14.8 g, 53.4 mmol), 2-chloro-4-(difluoromethyl)pyrimidine (8.79 g, 53.4 mmol), PdCl2 (xanthophos) (2.019 g, 2.67 mmol), and potassium carbonate (22.14 g, 160 mmol) were combined. The flask was sealed with a rubber diaphragm, and 1,4-dioxane (200 mL) and water (50.0 mL) were added to the flask. The flask was then refilled with argon (vacuumed, then refilled with argon three times). The mixture was stirred at 60°C for 3.5 hours. The mixture was cooled to room temperature, and volatile organic compounds were removed under reduced pressure to obtain an aqueous mixture. The slurry was taken into toluene (300 mL) and then further diluted with water (400 mL). The mixture was mixed and then filtered through a Celite pad to remove insoluble material. The organic layer was then separated, washed with brine, dried over MgSO4, filtered, and concentrated under reduced pressure. The residue was taken into a minimal amount of toluene (100 mL) and then mixed with Celite and concentrated under reduced pressure to obtain a free-flowing powder. This powder was divided into three equal parts, and each part was subjected to reverse-phase chromatography (415 g RediSep Gold C18 column) eluting (95:5 water:MeCN + 0.1% formic acid):(95:5 MeCN:water + 0.1% formic acid) 25:75 → 0:100. The fractions containing the desired product were pooled and partially concentrated under reduced pressure to obtain an aqueous mixture. The slurry was combined with toluene, and the aqueous layer was made slightly basic (pH 8) by adding 5N NaOH aqueous solution. The mixture was mixed, then the organic layer was isolated, washed with brine, dried over MgSO4, and concentrated under reduced pressure to obtain the product 2-amino-4-(4-(difluoromethyl)pyrimidine-2-yl)methyl benzoate (10.2 g, 68%) as a dark yellow solid.¹H NMR (500 MHz, CDCl₃) δ ppm 9.02 (br d, J=4.77 Hz, 1 H), 8.01 (d, J=8.34 Hz, 1 H), 7.86 (s, 1 H), 7.76 (br d, J=8.35 Hz, 1 H), 7.55 (br d, J=4.77 Hz, 1 H), 6.52 - 6.78 (m, 1 H), 5.88 (br s, 2 H), 3.94 (s, 3 H). LCMS method G: Retention time = 2.66 min; Observed ions = 321.1 (M + MeCN).

[0143] Step 3: Preparation of 2-amino-4-(4-(difluoromethyl)pyrimidine-2-yl)benzoic acid To a solution of 2-amino-4-(4-(difluoromethyl)pyrimidine-2-yl)methyl benzoate (12 g, 43.0 mmol) in methanol (50 mL) and THF (50.0 mL), 25.8 mL, 129 mmol, 5 N sodium hydroxide aqueous solution was added, and the mixture was stirred at 60°C for 1 hour. LC-MS analysis indicated that the reaction was complete. The mixture was cooled to room temperature, and then 1 M HCl aqueous solution (129 mL, 129 mmol) was added. To the dark yellow slurry, ELISA (250 mL) and water (150 mL) were added, and the yellow slurry partially dissolved. The organic layer was turbid, but the aqueous layer appeared homogeneous. The organic layer was washed with brine. The organic layer remained turbid. The organic layer was separated, and the turbid mixture was heated until it became a clear yellow solution. The solution was dried over MgSO4, filtered, and concentrated under reduced pressure to obtain the product 2-amino-4-(4-(difluoromethyl)pyrimidine-2-yl)benzoic acid (11.3 g, 99%) as a yellow solid. 1¹H NMR (500 MHz, CD3OD) δ ppm 9.06 (d, J=5.07 Hz, 1 H), 7.96 (d, J=8.35 Hz, 1 H), 7.93 (d, J=1.19 Hz, 1 H), 7.67 (dd, J=8.49, 1.64 Hz, 1 H), 7.64 (d, J=5.07 Hz, 1 H), 6.66 - 6.91 (m, 1 H). LC-MS method G: Retention time = 2.14 min; Observed ions = 307.0 (M + MeCN)

[0144] Another preparation of 2-amino-4-(4-(difluoromethyl)pyrimidine-2-yl)benzoic acid

[0145] [ka]

[0146] Synthesis scheme

[0147] [ka]

[0148] Preparation of methyl 4-bromo-2-nitrobenzoate This compound was prepared in 200g scale according to the procedure reported in WO2005037796 and J. Am. Chem. Soc., 2018, 140 (33), 10553-10561.

[0149] Step 1: Preparation of methyl 4-cyano-2-nitrobenzoate Under nitrogen at 27°C, copper(I) cyanide (234 g, 2615 mmol) was added to a stirred solution of methyl 4-bromo-2-nitrobenzoate (340 g, 1307 mmol) in DMF (3000 mL). The reaction mixture was then stirred at 150°C for 5 hours. The progress of the reaction was monitored by TLC (SiO2, 20% ethyl acetate / Pet., Rf=0.6, UV active). Once complete, the reaction mixture was cooled to 27°C. The reaction mixture was poured into ethyl acetate (5000 mL), and the resulting mixture was washed with a 5% aqueous ethylenediamine solution (5000 mL) to remove the copper salt. The organic solution was then washed with ice-cold water (3 × 3000 mL), followed by ice-cold brine (3000 mL). The organic layer was dried over Na2SO4, filtered, and concentrated under reduced pressure to obtain methyl 4-cyano-2-nitrobenzoate as a brown solid (280 g, 83%). The product was used directly in the next step without further purification. 1 HNMR (400 MHz, CDCl3) δ = 8.37 (d, J = 1.6 Hz, 1H), 7.97 (dd, J = 7.9, 1.6 Hz, 1H), 7.87 (d, J = 7.9 Hz, 1H), 3.97 (s, 3H). Note: The work-up process was further optimized to avoid the need for filtration. The reaction mixture was poured into ethyl acetate (5000 mL), and the resulting mixture was washed with a 5% aqueous ethylenediamine solution (5000 mL) to remove the copper salts. The organic solution was then washed with ice-cold water (3 × 3000 mL), followed by ice-cold brine (3000 mL). The remaining process is the same as described above.

[0150] Step 2: Preparation of 4-carbamidoyl-2-nitrobenzoate methyl hydrochloride Under nitrogen, sodium methoxide (44.0 g, 815 mmol) was added to a stirred solution of methyl 4-cyano-2-nitrobenzoate (280 g, 1358 mmol) in MeOH (4000 mL), and the reaction mixture was stirred at 27°C for 16 hours. Ammonium chloride (72.6 g, 1358 mmol) was added to the reaction mixture, and the reaction mixture was stirred at 27°C for 18 hours. The progress of the reaction was monitored by TLC (SiO2, 80% siRNA / Pet., Rf=0.1, UV active). Upon completion, the reaction mixture was filtered, and the filtered cake was extracted with a 10% MeOH DCM solution (3 × 1000 mL). The combined filtrate was concentrated under reduced pressure to obtain the crude product as a rubbery solid. This substance was triturated with siRNA (1000 mL) to obtain methyl 4-carbamimidol-2-nitrobenzoate hydrochloride as a yellow solid (250 g, 56%). The product was used directly in the next step without further purification. 1 H-NMR (400 MHz, DMSO-d6) δ = 9.01 (br s, 3H), 8.50 (d, J = 1.6 Hz, 1H), 8.23 ​​(dd, J = 7.9, 1.6 Hz, 1H), 8.08 (d, J = 7.9 Hz, 1H), 3.91 (s, 3H). LCMS purity = 79%.

[0151] Preparation of (E)-4-ethoxy-1,1-difluorobuta-3-en-2-one

[0152] [ka]

[0153] At 0°C, a mixture of pyridine (128 mL, 1580 mmol) and ethoxyethene (165 mL, 1724 mmol) was added dropwise over 1 hour to a stirred solution of 2,2-difluoroacetic anhydride (179 mL, 1436 mmol) in DCM (1250 mL). The reaction mixture was heated to 27°C and then stirred for 12 hours. The reaction mixture was quenched by adding ice-cold water (1000 mL). The organic layer was separated, washed with saturated NaHCO3 aqueous solution (1000 mL), then brine (1000 mL), dried over Na2SO4, and filtered. The filtrate was carefully concentrated under reduced pressure (pressure: ≥100 mbar; bath temperature: ≥25°C) to obtain (E)-4-ethoxy-1,1-difluorobuta-3-en-2-one as a brown liquid (180 g, 80%). The crude compound was used directly in the next step without further purification. 1 HNMR (400 MHz, CDCl3) δ = 7.84 (d, J = 12.4 Hz, 1H), 5.89-5.63 (m, 2H), 4.06 (q, J = 7.0 Hz, 2H), 1.39 (t, J = 7.0 Hz, 3H). GC-MS purity = 95%.

[0154] Step 3: Preparation of methyl 4-(4-(difluoromethyl)pyrimidine-2-yl)-2-nitrobenzoate Under a nitrogen atmosphere at 27°C, (E)-4-ethoxy-1,1-difluorobuta-3-en-2-one (159 g, 1062 mmol), followed by triethylamine (296 mL, 2124 mmol), was added to a 5 L autoclave flask containing a stirred solution of methyl 4-carbamimidyl-2-nitrobenzoate (200 g, 708 mmol) in EtOH (2000 mL). The reaction mixture was stirred at 80°C for 16 hours. The progress of the reaction was monitored by TLC (SiO2, 20% siRNA / Pet., Rf=0.5, UV active). The reaction mixture was filtered, and the filtrate was concentrated under reduced pressure to obtain methyl 4-(4-(difluoromethyl)pyrimidine-2-yl)-2-nitrobenzoate as a brown liquid (220 g, 58%). The product was used directly in the next step without further purification. 1H-NMR (400 MHz, CDCl3) δ = 9.07-9.00 (m, 1H), 8.80 (dd, J = 7.9, 1.6 Hz, 1H), 7.88 (d, J = 7.9 Hz, 1H), 7.78-7.56 (m, 1H), 7.62 (d, J = 4.8 Hz, 1H), 6.65 (t, J = 54.8 Hz, 1H), 3.96 (s, 3H). HPLC Purity: 58%.

[0155] Step 4: Preparation of 2-amino-4-(4-(difluoromethyl)pyrimidine-2-yl)methyl benzoate At 27°C, methyl 4-(4-(difluoromethyl)pyrimidine-2-yl)-2-nitrobenzoate (220 g, 711 mmol) was mixed with EtOH (2150 mL) and water (215 mL) and then ammonium chloride (190 g, 3557 mmol) was added, followed by iron (199 g, 3557 mmol). The reaction mixture was stirred at 80°C for 16 hours. The progress of the reaction was monitored by TLC (SiO2, 20% siRNA / Pet., Rf=0.4, UV active). Upon completion, the reaction mixture was filtered while still hot through a Celite pad, and the Celite pad was extracted with siRNA (4 × 500 mL). The combined filtrate was concentrated under reduced pressure to obtain methyl 2-amino-4-(4-(difluoromethyl)pyrimidine-2-yl)benzoate as a yellow solid (230 g, 64%). The product was used directly in the next step without further purification. 1 H-NMR (400 MHz, CDCl3) δ = 8.99 (d, J = 4.8 Hz, 1H), 7.98 (d, J = 8.3 Hz, 1H), 7.83 (s, 1H), 7.73 (d, J = 8.4 Hz, 1H), 7.52 (d, J = 4.8 Hz, 1H), 6.61 (t, J = 54.8 Hz, 1H), 3.91 (s, 3H). LCMS purity = 56%.

[0156] Step 5: Preparation of 2-amino-4-(4-(difluoromethyl)pyrimidine-2-yl)benzoic acid At 27°C, 230 g (461 mmol) of methyl 2-amino-4-(4-(difluoromethyl)pyrimidine-2-yl)benzoate was mixed with 2300 mL of THF, 575 mL of MeOH, and 192 mL of water. LiOH (66.3 g, 2767 mmol) was added to this mixed solution. The reaction mixture was stirred at 50°C for 16 hours. The progress of the reaction was monitored by TLC (SiO2, 40% HCl / Pet., Rf=0.1, UV active). Once complete, the reaction mixture was cooled to 27°C and then concentrated under reduced pressure. The crude residue was dissolved in water (1000 mL) and washed with HCl (2 × 250 mL). The aqueous layer was acidified to approximately pH 6 with 1N HCl aqueous solution. The precipitated solid was collected by filtration, washed with water (500 mL), then with n-pentane (500 mL), and then dried to obtain methyl 2-amino-4-(4-(difluoromethyl)pyrimidine-2-yl)benzoate as a yellow solid (80 g, 63%). 1 H-NMR (400 MHz, DMSO-d6) δ = 9.15 (d, J = 4.8 Hz, 1H), 7.87-7.84 (m, 2H), 7.74 (d, J = 4.8 Hz, 1H), 7.53 (d, J = 8.4 Hz, 1H), 7.06 (t, J = 54.2 Hz, 1H). LCMS purity = 96%.

[0157] Example 2: Preparation of N-((S)-1-((3P)-3-(4-chloro-1-methyl-3-(methylsulfonamide)-1H-indazole-7-yl)-7-(4-(difluoromethyl)pyrimidine-2-yl)-4-oxo-3,4-dihydroquinazoline-2-yl)-2-(3,5-difluorophenyl)ethyl)-2-((3bS,4aR)-3-(difluoromethyl)-5,5-difluoro-3b,4,4a,5-tetrahydro-1H-cyclopropa[3,4]cyclopenta[1,2-c]pyrazole-1-yl)acetamide

[0158] [ka]

[0159] Synthesis scheme

[0160] [ka]

[0161] Step 1: Preparation of tert-butyl(S)-(1-(3-(4-chloro-3-(N-(4-methoxybenzyl)methylsulfonamide)-1-methyl-1H-indazole-7-yl)-7-(4-(difluoromethyl)pyrimidine-2-yl)-4-oxo-3,4-dihydroquinazoline-2-yl)-2-(3,5-difluorophenyl)ethyl)carbamate (S)-2-((tert-butoxycarbonyl)amino)-3-(3,5-difluorophenyl)propanoic acid (76 g, 253 mmol) and 2-amino-4-(4-(difluoromethyl)pyrimidine-2-yl)benzoic acid (73.9 g, 279 mmol) were mixed in acetonitrile (2.1 L) and pyridine (0.049 L, 608 mmol) was added. The reaction mixture was cooled to -5°C and stirred at the same temperature for 10 minutes. Then, T3P (50% ethyl acetate solution, 0.754 L, 1266 mmol) was slowly added to the reaction mixture at -5°C. The mixture was stirred at -5°C for 20 minutes, then heated to 27°C and stirred for 2 hours. N-(7-amino-4-chloro-1-methyl-1H-indazole-3-yl)-N-(4-methoxybenzyl)methanesulfonamide (100 g, 253 mmol) was added to the reaction mixture all at once at 27°C, and the mixture was stirred for 18 hours. The progress of the reaction was monitored by TLC (SiO2, 40% siRNA / PET, Rf=0.4, UV active). The reaction mixture was concentrated under reduced pressure to remove acetonitrile, then diluted with siRNA (1000 mL) and washed with water (2000 mL). The organic layer was separated, washed with saturated Na2SO3 aqueous solution (3 × 500 mL), then brine (500 mL), dried over anhydrous Na2SO4, filtered, and concentrated under reduced pressure to obtain the crude product as a brown, rubbery liquid. This was purified by column chromatography on silica gel eluted with 30-40% siRNA / PET. The fraction containing the desired product was collected and concentrated under reduced pressure to obtain tert-butyl(S)-(1-(3-(4-chloro-3-(N-(4-methoxybenzyl)methylsulfonamide)-1-methyl-1H-indazole-7-yl)-7-(4-(difluoromethyl)pyrimidine-2-yl)-4-oxo-3,4-dihydroquinazoline-2-yl)-2-(3,5-difluorophenyl)ethyl)carbamate (180 g, 78%, yellow solid) as a mixture of homochiral atropisomers (diastereomers).

[0162] Step 2: Preparation of (S)-N-((6P)-7-(2-(1-amino-2-(3,5-difluorophenyl)ethyl)-7-(4-(difluoromethyl)pyrimidine-2-yl)-4-oxoquinazoline-3(4H)-yl)-4-chloro-1-methyl-1H-indazole-3-yl)methanesulfonamide Under an N2 atmosphere at 27°C, tert-butyl(S)-(1-(3-(4-chloro-3-(N-(4-methoxybenzyl)methylsulfonamide)-1-methyl-1H-indazole-7-yl)-7-(4-(difluoromethyl)pyrimidine-2-yl)-4-oxo-3,4-dihydroquinazolin-2-yl)-2-(3,5-difluorophenyl)ethyl)carbamate (110 g, 121 mmol) was stirred in DCM (500 mL), to which TFA (374 mL, 4849 mmol) was added, and the solution was stirred for 10 minutes. Trifluoromethanesulfonic acid (32.3 mL, 364 mmol) was added to the solution, and the solution was stirred at 27°C for 1 hour. The progress of the reaction was monitored by TLC (SiO2, 50% Â / Pet., Rf=0.2). Volatile substances were removed under a gentle stream of nitrogen gas. The resulting residue was dissolved in 1500 mL of ethyl acetate, then washed with 2 × 750 mL of 1 M aqueous NaOH solution, then with brine (750 mL), dried over Na₂SO₄, filtered, and concentrated under reduced pressure to obtain the crude product as an off-white solid. This substance was purified by silica gel chromatography eluting with 80–98% ethyl acetate / PET. The fraction containing the desired product was recovered and concentrated under reduced pressure to obtain (S)-N-(7-(2-(1-amino-2-(3,5-difluorophenyl)ethyl)-7-(4-(difluoromethyl)pyrimidine-2-yl)-4-oxoquinazoline-3(4H)-yl)-4-chloro-1-methyl-1H-indazole-3-yl)methanesulfonamide as a yellow solid (65 g, 74%). The product is a mixture of homochiral atropisomers (diastereomers). The above procedure was repeated four more times to obtain a total of 310 g of (S)-N-(7-(2-(1-amino-2-(3,5-difluorophenyl)ethyl)-7-(4-(difluoromethyl)pyrimidine-2-yl)-4-oxoquinazoline-3(4H)-yl)-4-chloro-1-methyl-1H-indazole-3-yl)methanesulfonamide, which was dissolved in DCM:MeCN (30:70, 3055 mL).Next, the product was purified by preparative SFC using the following method (column = (R,R)Welk-01, 30 × 250 mm, 5 μm, eluent = CO2:methanol (1:1); flow rate = 90.0 g / min; back pressure = 120.0 bar; detection = 254 nm (UV); stack time = 16.0 min; input per injection = 800 mg). Separation produced two peaks. The major peak (second to elute) was collected and concentrated under reduced pressure to obtain (S)-N-((6P)-7-(2-(1-amino-2-(3,5-difluorophenyl)ethyl)-7-(4-(difluoromethyl)pyrimidine-2-yl)-4-oxoquinazoline-3(4H)-yl)-4-chloro-1-methyl-1H-indazole-3-yl)methanesulfonamide as a yellow solid (170 g, 51%). The product is a single stereoisomer. 1 H NMR (400 MHz, CDCl3) δ = 9.16-9.07 (m, 1H), 9.01-8.95 (m, 1H), 8.71-8.62 (m, 1H), 8.46-8.37 (m, 1H), 7.68-7.61 (m, 1H), 7.11 (d, J = 7.8 Hz, 1H), 6.86-6.55 (m, 2H), 6.54-6.45 (m, 3H), 3.79-3.74 (m, 3H), 3.71-3.63 (m, 1H), 3.44-3.33 (m, 4H), 2.94-2.83 (m, 1H). LCMS Purity = 94%.

[0163] Step 3: Preparation of N-((S)-1-((3P)-3-(4-chloro-1-methyl-3-(methylsulfonamide)-1H-indazole-7-yl)-7-(4-(difluoromethyl)pyrimidine-2-yl)-4-oxo-3,4-dihydroquinazoline-2-yl)-2-(3,5-difluorophenyl)ethyl)-2-((3bS,4aR)-3-(difluoromethyl)-5,5-difluoro-3b,4,4a,5-tetrahydro-1H-cyclopropa[3,4]cyclopenta[1,2-c]pyrazole-1-yl)acetamide At 27℃, (S)-N-((6P)-7-(2-(1-amino-2-(3,5-difluorophenyl)ethyl)-7-(4-(difluoromethyl)pyrimidine-2-yl)-4-oxoquinazoline-3(4H)-yl)-4-chloro-1-methyl-1H-indazole-3-yl)methanesulfonamide (50g, 61.9mmol), 2-((3bS,4aR)-3-(difluoromethyl)-5,5-difluoro-3b,4,4a,5-tetrahydro- To a solution of 1H-cyclopropa[3,4]cyclopenta[1,2-c]pyrazole-1-yl)acetic acid (16.34 g, 61.9 mmol) and 1-hydroxybenzotriazole hydrate ("HOBt hydrate", 3.79 g, 24.74 mmol) in DMF (500 mL), N-methylmorpholine (13.60 mL, 124 mmol) and N-(3-dimethylaminopropyl)-N'-ethylcarbodiimide hydrochloride (21.34 g, 111 mmol) were added. The reaction mixture was stirred at 27 °C for 16 hours. The progress of the reaction was monitored by TLC (SiO2, 50% SiO2 / Pet., Rf=0.5, UV active). The reaction mixture was diluted with ice-cold water (7 L) and then stirred for 30 minutes. The precipitated solid was collected by filtration and then dried under vacuum to obtain the crude compound as an off-white solid (75 g). LCMS purity = 60%.The above procedure was repeated three more times to obtain a total of 185 g of crude N-((S)-1-((3P)-3-(4-chloro-1-methyl-3-(methylsulfonamide)-1H-indazole-7-yl)-7-(4-(difluoromethyl)pyrimidine-2-yl)-4-oxo-3,4-dihydroquinazolin-2-yl)-2-(3,5-difluorophenyl)ethyl)-2-((3bS,4aR)-3-(difluoromethyl)-5,5-difluoro-3b,4,4a,5-tetrahydro-1H-cyclopropa[3,4]cyclopenta[1,2-c]pyrazole-1-yl)acetamide, which was then blended and purified by silica gel chromatography eluted with 30-40% HCl / Pet. The fraction containing the desired product was collected and concentrated under reduced pressure to obtain the desired product as an off-white solid (100 g, LCMS purity: 97%). This substance was suspended in isopropanol (1000 mL, 10V), heated at 70°C for 30 minutes, and then slowly cooled to 27°C over 16 hours to obtain the crystalline product. The obtained solid was recovered by filtration and then dried under vacuum to obtain N-((S)-1-((3P)-3-(4-chloro-1-methyl-3-(methylsulfonamide)-1H-indazole-7-yl)-7-(4-(difluoromethyl)pyrimidine-2-yl)-4-oxo-3,4-dihydroquinazolin-2-yl)-2-(3,5-difluorophenyl)ethyl)-2-((3bS,4aR)-3-(difluoromethyl)-5,5-difluoro-3b,4,4a,5-tetrahydro-1H-cyclopropa[3,4]cyclopenta[1,2-c]pyrazole-1-yl)acetamide as an off-white solid (80 g, 79%).1H NMR (acetone-d6) δ: 9.27 (d, J=5.1 Hz, 1H), 8.89 (d, J=1.8 Hz, 1H), 8.69 (dd, J=8.3, 1.8 Hz, 1H), 8.57 (br s, 1H), 8.39 (d, J=8.3 Hz, 1H), 8.12 (d, J=8.9 Hz, 1H), 7.84 (d, J=5.1 Hz, 1H), 7.51 (d, J=7.7 Hz, 1H), 7.37 (d, J=8.0 Hz, 1H), 7.03 (t, J=54.4 Hz, 1H), 6.86 (tt, J=9.2, 2.4 Hz, 1H), 6.70-6.76 (m, 2H), 6.78 (t, J=54.7 Hz, 1H), 4.93 (td, J=9.0, 4.6 Hz, 1H), 4.65-4.76 (m, 2H), 3.69 (s, 3H), 3.56 (dd, J=14.2, 4.6 Hz, 1H), 3.27 (s, 3H), 3.15 (dd, J=14.3, 9.2 Hz, 1H), 2.42-2.53 (m, 2H), 1.37-1.44 (m, 1H), 0.95-1.00 (m, 1H).LCMS method: Column = Acquity BEH C18, 2.1×50mm, 1.7μm Particles; Solvent A = 0.1% formic acid aqueous solution; Solvent B = 0.1% formic acid in acetonitrile solution; Flow rate = 0.6 mL / min; Gradient {Time point (min) / %B (%) at time point} = 0 / 3, 0.4 / 3, 3.2 / 98, 3.8 / 98, 4.2 / 3, 4.5 / 3; Column temperature = 35°C. LCMS results: Retention time = 2.73 min; Observed ions = 933.09 (M + H); Purity = 99%.

[0164] Naming of Example 2: The compound of Example 2 prepared above is a homochiral substance containing axial chirality. Axial chirality can be described using the O / M nomenclature detailed in the IUPAC Gold Book (doi:10.1351 / goldbook.A00547). However, at present, the number of software tools capable of generating chemical names including P / M nomenclature is limited, and the options for converting chemical names using this nomenclature into molecular structural representations are even more limited. Therefore, for clarity and convenience, some of the names of Example 2 are listed below.

[0165] The names of Example 2, generated by ChemDraw Professional 16 (without P / M nomenclature), are as follows: N-((S)-1-(3-(4-chloro-1-methyl-3-(methylsulfonamide)-1H-indazole-7-yl)-7-(4-(difluoromethyl)pyrimidine-2-yl)-4-oxo-3,4-dihydroquinazoline-2-yl)-2-(3,5-difluorophenyl)ethyl)-2-((3bS,4aR)-3-(difluoromethyl)-5,5-difluoro-3b,4,4a,5-tetrahydro-1H-cyclopropa[3,4]cyclopenta[1,2-c]pyrazole-1-yl)acetamide

[0166] The chemical names of Example 2, generated by JChem for Excel (including P / M nomenclature), are as follows: N-[(1S)-1-[(3P)-3-(4-chloro-3-methanesulfonamide-1-methyl-1H-indazole-7-yl)-7-[4-(difluoromethyl)pyrimidine-2-yl]-4-oxo-3,4-dihydroquinazoline-2-yl]-2-(3,5-difluorophenyl)ethyl]-2-[(2S,4R)-9-(difluoromethyl)-5,5-difluoro-7,8-diazatricyclo[4.3.0.0 2 , 4 ]nona-1(6),8-dien-7-yl]acetamide

[0167] The chemical names for Example 2, generated by ChemDraw Professional 16 with manual addition of P / M nomenclature, are as follows: N-((S)-1-((3P)-3-(4-chloro-1-methyl-3-(methylsulfonamide)-1H-indazole-7-yl)-7-(4-(difluoromethyl)pyrimidine-2-yl)-4-oxo-3,4-dihydroquinazoline-2-yl)-2-(3,5-difluorophenyl)ethyl)-2-((3bS,4aR)-3-(difluoromethyl)-5,5-difluoro-3b,4,4a,5-tetrahydro-1H-cyclopropa[3,4]cyclopenta[1,2-c]pyrazole-1-yl)acetamide

[0168] Preparation of 3,3-difluorobutan-1-ol

[0169] [ka]

[0170] Synthesis scheme [ka]

[0171] Step 1: Preparation of 3-oxobutylbenzoate

[0172] [ka]

[0173] Under a nitrogen atmosphere at -70°C, pyridine (470 mL) was added dropwise over 1 hour to a stirred solution of benzoyl chloride (0.396 L, 3405 mmol) in DCM (1 L). After stirring at the same temperature for 30 minutes, a solution of 4-hydroxybutan-2-one (250.0 g, 2837 mmol) in DCM (500 mL) was added dropwise over 1 hour. The reaction mixture was heated to 26°C and then stirred for 16 hours. The progress of the reaction was monitored by TLC (SiO2, 30% Â / Pet. Rf=0.4). Upon completion, the reaction mixture was washed with water (2 × 1000 mL), 1N HCl (2 × 500 mL), and then saturated NaHCO3 solution (2 × 500 mL). The organic layer was dried over Na2SO4, filtered, and concentrated under reduced pressure to obtain 3-oxobutylbenzoate as a pale yellow liquid (400 g, yield = 69%). 1 H NMR (400 MHz, CHLOROFORM-d) δ = 8.05 - 7.94 (m, 2H), 7.60 - 7.51 (m, 1H), 7.47 - 7.36 (m, 2H), 4.65 - 4.54 (t, 2H), 2.97 - 2.84 (t, 2H), 2.28 - 2.13 (s, 3H). HPLC purity = 94.1%.

[0174] Step 2: Preparation of 3,3-difluorobutylbenzoate

[0175] [ka]

[0176] Under a nitrogen atmosphere at 0°C, DAST (677 mL, 5125 mmol) was added dropwise over 1 hour to a stirred solution of 3-oxobutylbenzoate (90 g, 427 mmol) in dichloromethane (700 mL). The reaction mixture was heated to 26°C and stirred for 16 hours. The reaction was monitored by TLC (SiO2, 20% Â / PET. Rf=0.6). Once complete, the reaction mixture was diluted with DCM (500 mL) and slowly poured into a cold saturated NaHCO3 (1 L) aqueous solution. The organic layer was separated, washed with brine solution (400 mL), dried over anhydrous Na2SO4, filtered, and concentrated under reduced pressure to obtain the crude compound (95 g) as a yellow liquid. This substance was purified by column chromatography using silica gel (100-200 mesh) and eluted with a 0-5% Â PET. solution. The fraction containing the product was collected and concentrated under reduced pressure to obtain 3,3-difluorobutylbenzoate (60 g, yield=59%) as a brown liquid. 1 H NMR (400 MHz, CDCl3) δ = 8.06 - 8.01 (m, 2H), 7.60 - 7.54 (m, 1H), 7.48 - 7.40 (m, 2H), 4.54 - 4.48 (t, 2H), 2.43 - 2.29 (m, 2H), 1.77 - 1.64 (m, 3H). LCMS purity = 89.74%; m / z = 215.33.

[0177] Step 3: Preparation of 3,3-difluorobutan-1-ol

[0178] [ka]

[0179] Under a nitrogen atmosphere at 0°C, a solution of 3,3-difluorobutylbenzoate (100 g, 467 mmol) in THF (800 mL) was stirred, to which a solution of lithium hydroxide monohydrate (137 g, 3268 mmol) in water (800 mL) was added. The reaction mixture was heated to 26°C and then stirred for 16 hours. The progress of the reaction was monitored by TLC (SiO2, 20% siRNA / Pet. Rf=0.6, KMnO4 active). Upon completion, the reaction mixture was diluted with diethyl ether (400 mL). The organic layer was separated, and the aqueous layer was re-extracted with diethyl ether (300 mL). The combined organic compounds were washed with brine (200 mL), dried over Na2SO4, filtered, and concentrated under reduced pressure (volatile products, bath temperature = 25°C) to obtain the crude compound as a black liquid. This substance was diluted with diethyl ether (100 mL) and treated with charcoal. The mixture was filtered through a Celite pad. The Celite pad was extracted with diethyl ether (200 mL). The combined filtrate was concentrated under reduced pressure (volatile product, bath temperature = 25°C) to obtain 3,3-difluorobutan-1-ol (40 g, yield = 71%) as a pale yellow liquid. 1 H-NMR (400 MHz, CDCl3) δ = 3.87 (t, J = 6.1 Hz, 2H), 2.22 - 2.07 (m, 2H), 1.73 - 1.57 (m, 3H). GCMS Purity = 91.3%; m / z = 110.0.

[0180] Preparation of 2-amino-6-(benzyloxy)nicotinic acid [ka]

[0181] Synthesis scheme [ka]

[0182] Step 1: Preparation of 2-amino-6-(benzyloxy)nicotinic acid

[0183] [ka]

[0184] Under an N2 atmosphere at 26°C, potassium tert-butoxide (390 g, 3477 mmol) was added to a stirred solution of 2-amino-6-chloronicotinic acid (200 g, 1159 mmol) in benzyl alcohol (1400 mL, 13464 mmol). The reaction mixture was heated to 120°C and stirred at that temperature for 16 hours. The progress of the reaction was monitored by TLC (SiO2, 10% MeOH DCM solution, Rf=0.5). Upon completion, the reaction mixture was diluted with water (3 L) and extracted with diethyl ether (2 × 1000 mL). The organic layer was separated, and the aqueous layer was acidified to pH 4 using aqueous citric acid (0.5 M). The precipitated solid was recovered by filtration and then dried under reduced pressure to obtain 2-amino-6-(benzyloxy)nicotinic acid (220 g, yield = 72%) as an off-white solid. 1 H NMR (400 MHz, DMSO-d6) δ = 12.56 - 12.32 (m, 1H), 7.97 - 7.91 (m, 1H), 7.52 - 7.41 (m, 2H), 7.38 - 7.11 (m, 5H), 6.03 (d, J = 8.5 Hz, 1H), 5.39 - 5.31 (m, 2H). LCMS Purity = 93%; m / z = 245.29 (M+H).

[0185] Step 2: Preparation of methyl 2-amino-6-(benzyloxy)nicotinate

[0186] [ka]

[0187] Under an N2 atmosphere at 26°C, potassium carbonate (373 g, 2702 mmol) and iodomethane (0.282 L, 4504 mmol) were slowly added to a stirred solution of 2-amino-6-(benzyloxy)nicotinic acid (220 g, 901 mmol) in DMF (2.5 L). The reaction mixture was stirred at 27°C for 16 hours. The progress of the reaction was monitored by TLC (SiO2, 40% siRNA / Pet., Rf=0.6). Upon completion, the reaction mixture was diluted with water (5 L). The precipitated solid was isolated by filtration and then dried under vacuum to obtain methyl 2-amino-6-(benzyloxy)nicotinate (220 g, yield=92%) as an off-white solid. 1 H NMR (400 MHz, CDCl3) δ = 8.00 (d, J = 8.4 Hz, 1H), 7.42-7.40 (m, 2H), 7.39-7.35 (m, 2H), 7.34-7.31 (m, 1H), 6.01 (d, J = 8.4 Hz, 1H), 5.33 (s, 2H), 3.84 (s, 3H). LCMS Purity = 97%, m / z = 259.30 (M+H).

[0188] Step 3: Preparation of methyl 2-amino-6-hydroxynicotinate

[0189] [ka]

[0190] Under an N2 atmosphere at 26°C, 800 mL of TFA and 25 mL of trifluoromethanesulfonic acid (282 mmol) were slowly added to a stirred solution of methyl 2-amino-6-(benzyloxy)nicotinate (50 g, 190 mmol) in DCM (500 mL). The reaction mixture was stirred at 26°C for 16 hours. The progress of the reaction was monitored by TLC (SiO2, siRNA, Rf=0.2). Upon completion, volatile substances were removed under vacuum to obtain the crude product. This product was triturated with diethyl ether (3 × 1000 mL), and the precipitated solid was isolated by filtration. Water (2 L) was added to the solid, and the mixture was stirred for 5 hours. The solid was recovered by filtration and washed with water. The solid was dried under vacuum to obtain methyl 2-amino-6-hydroxynicotinate (25 g, yield = 78%) as an off-white solid. ¹H NMR (300 MHz, DMSO-d6) δ = 10.92-10.76 (m, ¹H), 7.65 (d, J = 9.5 Hz, ¹H), 7.43-6.87 (m, ²H), 5.51 (d, J = 9.5 Hz, ¹H), 3.69 (s, ³H). LCMS Purity = 99.32%; m / z = 169.32 (M+H). The absence of TFA and trifluoromethanesulfonic acid in the product was confirmed. 19 Confirmation was made by 1F-NMR. The product was used directly in the next step without further purification.

[0191] Step 4: Preparation of methyl 2-amino-6-(3,3-difluorobutoxy)nicotinate

[0192] [ka]

[0193] Under an N2 atmosphere at 0°C, triphenylphosphine (77 g, 294 mmol) was added to a stirred solution of methyl 2-amino-6-hydroxynicotinate (25 g, 147 mmol) in THF (375 mL), followed by the dropwise addition of DIAD (57.2 mL, 294 mmol). The reaction mixture was stirred at 0°C for 15 minutes, and then a solution of 3,3-difluorobutan-1-ol (25.3 g, 221 mmol) in THF (125 mL) was added dropwise at 0°C. The reaction mixture was brought to 27°C and then stirred for 5 hours. The progress of the reaction was monitored by TLC (SiO2, siRNA, Rf=0.5). Upon completion, the reaction mixture was concentrated under reduced pressure to obtain the crude product. This substance was stirred in MTBE:pet. (1:1, 1 L). The mixture was filtered, and the filter pad was extracted with MTBE:pet. (1:1, 4 × 200 mL). The combined filtrate was concentrated under reduced pressure to obtain a pale yellow, rubbery solid. This substance was purified by column chromatography using silica gel (100-200 mesh) and eluted with a 10-20% siRNA solution in petroleum. The fraction containing the product was collected and concentrated under reduced pressure to obtain methyl 2-amino-6-(3,3-difluorobutoxy)nicotinate (20 g, yield = 48%) as a pale yellow liquid. 1 H NMR (400 MHz, CDCl3) δ = 8.05 - 7.95 (m, 1H), 6.02 (d, J = 8.8 Hz, 1H), 4.45 (t, J = 6.8 Hz, 2H), 3.80 (s, 3H), 2.40 - 2.22 (m, 2H), 1.68 (t, J = 18.6 Hz, 3H). LCMS purity = 91.1%, m / z = 261.25 (M+H).

[0194] Step 5: Preparation of 2-amino-6-(3,3-difluorobutoxy)nicotinic acid

[0195] [ka]

[0196] At 26 °C, an aqueous solution of LiOH (2.491 g, 104 mmol) in water (30 mL) was added to a stirred solution of methyl 2-amino-6-(3,3-difluorobutoxy)nicotinate (5.7 g, 20.81 mmol) in THF (120 mL) and methanol (30 mL). The reaction mixture was heated to 70 °C and stirred at that temperature for 16 h. The progress of the reaction was monitored by TLC (SiO2, 50% EtOAc / Pet Rf = 0.2). Upon completion, the reaction mixture was concentrated under reduced pressure. The resulting residue was dissolved in water (60 mL) and acidified to pH 4 using 1N HCl. The mixture was extracted with ethyl acetate (3 × 100 mL). The combined organics were washed with brine (100 mL), dried over anhydrous Na2SO4, and concentrated under reduced pressure to give 2-amino-6-(3,3-difluorobutoxy)nicotinic acid (4.6 g, yield = 87%) as a brown solid. 1 1H NMR (400 MHz, CDCl3) δ = 11.66 - 10.84 (m, 1H), 8.12 - 7.97 (m, 1H), 6.07 (d, J = 8.3 Hz, 1H), 4.52 - 4.36 (m, 2H), 2.41 - 2.28 (m, 2H), 1.68 (t, J = 18.6 Hz, 3H). LCMS Purity = 97.68%, m / z = 247.24 (M+H).

[0197] Example 3: Preparation of N-((S)-1-((3P)-3-(4-chloro-1-methyl-3-(methylsulfonamide)-1H-indazole-7-yl)-7-(3,3-difluorobutoxy)-4-oxo-3,4-dihydropyrido[2,3-d]pyrimidine-2-yl)-2-(3,5-difluorophenyl)ethyl)-2-((3bS,4aR)-3-(difluoromethyl)-5,5-difluoro-3b,4,4a,5-tetrahydro-1H-cyclopropa[3,4]cyclopenta[1,2-c]pyrazole-1-yl)acetamide

[0198]

Chem.

[0199] Synthesis Scheme

Chem.

[0200] Step 1: Preparation of tert-butyl(S)-(1-(3-(4-chloro-3-(N-(4-methoxybenzyl)methylsulfonamide)-1-methyl-1H-indazole-7-yl)-7-(3,3-difluorobutoxy)-4-oxo-3,4-dihydropyrido[2,3-d]pyrimidine-2-yl)-2-(3,5-difluorophenyl)ethyl)carbamate

[0201] [ka]

[0202] Under an N2 atmosphere at -25°C, pyridine (47.0 mL, 581 mmol) was added to a stirred solution of (S)-2-((tert-butoxycarbonyl)amino)-3-(3,5-difluorophenyl)propanoic acid (50 g, 166 mmol) and 2-amino-6-(3,3-difluorobutoxy)nicotinic acid (41.3 g, 166 mmol) in acetonitrile (1000 mL). To the resulting mixture, 2,4,6-tripropyl-1,3,5,2,4,6-trioxatriphosfinan 2,4,6-trioxide ("T3P", 50 wt% ethyl ethyl solution, 494 mL, 830 mmol) was added dropwise over 15 minutes. The solution was heated to 13°C and then stirred for 5 hours. N-(7-amino-4-chloro-1-methyl-1H-indazole-3-yl)-N-(4-methoxybenzyl)methanesulfonamide (62.3 g, 158 mmol) was added to a solution at 13°C. The reaction mixture was then slowly heated to 27°C and stirred at that temperature for 48 hours. The progress of the reaction was monitored by TLC (SiO2, 50% siRNA / Pet., Rf=0.4). Upon completion, the reaction mixture was concentrated under reduced pressure, and the residue was added dropwise to saturated NaHCO3 aqueous solution (1000 mL) at 0°C. A white precipitate formed and was collected by vacuum filtration. The isolated solid was washed with water (2 L). Vacuum filtration was maintained until most of the residual water was removed from the solid. The solid was then dissolved in DCM (2 L). The solution was dried over Na2SO4, filtered, and then concentrated under reduced pressure to obtain the crude product. The substance was purified by silica gel chromatography eluting with a 50-65% ethyl acetate solution in PET. The fraction containing the desired product was pooled and concentrated under reduced pressure to obtain tert-butyl(S)-(1-(3-(4-chloro-3-(N-(4-methoxybenzyl)methylsulfonamide)-1-methyl-1H-indazole-7-yl)-7-(3,3-difluorobutoxy)-4-oxo-3,4-dihydropyrido[2,3-d]pyrimidine-2-yl)-2-(3,5-difluorophenyl)ethyl)carbamate (50 g, yield = 31%) as a yellow foamy solid. The above procedure was repeated seven more times on the same scale to produce a total of 592 g of product. The combined product (592 g) was dissolved in MeOH (1 L).The solution was diluted with n-hexane (6 L). An off-white solid precipitated, and the suspension was then stirred for 20 minutes. The solid was recovered by vacuum filtration, and the filtrate was secured. The solid was dried under vacuum to obtain tert-butyl(S)-(1-(3-(4-chloro-3-(N-(4-methoxybenzyl)methylsulfonamide)-1-methyl-1H-indazole-7-yl)-7-(3,3-difluorobutoxy)-4-oxo-3,4-dihydropyrido[2,3-d]pyrimidine-2-yl)-2-(3,5-difluorophenyl)ethyl)carbamate (300 g, yield = 48%) as an off-white solid. This product is a mixture of homochiral atropisomers (diastereomers). LCMS analysis method: Column = X Bridge BEH C18 (50 mm × 4.6 mm, 2.5 μm particles); Mobile phase A = 5 mM ammonium bicarbonate; Mobile phase B = acetonitrile; Gradient profile (time (min) / %B) = 0 / 5, 0.5 / 5, 1.5 / 15, 7 / 98, 9 / 98, 9.5 / 5, 10 / 5; Column temperature = 35°C; Flow rate = 1.3 mL / min. LCMS results: Retention time = 6.20 min. Observed ions = 888.09 (M + H); LCMS purity = 95%. Note: The retained filtrate was concentrated and dried under vacuum to obtain the product (120 g, pale yellow solid), which was also used separately from the above product in downstream chemistry.

[0203] Step 2: Preparation of (S)-N-((6P)-7-(2-(1-amino-2-(3,5-difluorophenyl)ethyl)-7-(3,3-difluorobutoxy)-4-oxopyrid[2,3-d]pyrimidine-3(4H)-yl)-4-chloro-1-methyl-1H-indazole-3-yl)methanesulfonamide

[0204] [ka]

[0205] To a stirred solution of tert-butyl(S)-(1-(3-(4-chloro-3-(N-(4-methoxybenzyl)methylsulfonamide)-1-methyl-1H-indazole-7-yl)-7-(3,3-difluorobutoxy)-4-oxo-3,4-dihydropyrido[2,3-d]pyrimidine-2-yl)-2-(3,5-difluorophenyl)ethyl)carbamate (95% purity, 300 g, 321 mmol) in DCM (3000 mL), trifluoroacetic acid (TFA) (900 mL) and then trifluoromethanesulfonic acid (158 mL, 1782 mmol) were added at 0°C. The solution was heated to 27°C and then stirred under a nitrogen atmosphere for 2 hours. The progress of the reaction was monitored by TLC (SiO2, 80% Â / Pet. Rf=0.3). Upon completion, volatile substances were removed under a gentle stream of nitrogen gas. The residue was added to a saturated NaHCO3 solution (1000 mL) at 0°C. The pH of the solution was adjusted to approximately 8 by adding solid NaHCO3. The mixture was extracted with Depositphotos (5 × 1000 mL). The combined organic layer was dried over Na2SO4, filtered, and then concentrated under reduced pressure to obtain the crude product. This product was purified by silica gel chromatography eluting with a 5–10% MeOH DCM solution. The fraction containing the desired product was pooled and concentrated under reduced pressure to obtain (S)-N-(7-(2-(1-amino-2-(3,5-difluorophenyl)ethyl)-7-(3,3)-difluorobutoxy)-4-oxopyrido[2,3-d]pyrimidine-3(4H)-yl)-4-chloro-1-methyl-1H-indazole-3-yl)methanesulfonamide (211 g, brown foamy solid) as a mixture of homochiral atropisomers (diastereomer, major: 79%, minor: 10% by LC-MS). This substance was dissolved in methanol:acetonitrile (80:20, 1800 mL) and purified by preparative SFC using the following method (column = (R,R)WHELK-01 (30 × 250 mm, 5 μm particles), eluent = CO2:MeOH (60:40); flow rate = 90 g / min; back pressure = 100 bar; detection = 214 nm (UV); stack time = 15.5 min; input per injection = 1.125 g).The pure major peak was collected and concentrated under reduced pressure to obtain (S)-N-((6P)-7-(2-(1-amino-2-(3,5-difluorophenyl)ethyl)-7-(3,3-difluorobutoxy)-4-oxopyrid[2,3-d]pyrimidine-3(4H)-yl)-4-chloro-1-methyl-1H-indazole-3-yl)methanesulfonamide (151 g, yield = 69%) as a brown solid. The product is a single stereoisomer. 1H-NMR (400 MHz, DMSO-d6) δ: 8.41 (d, J = 8.8 Hz, 1H), 7.39 (dd, J = 22.4, 7.9 Hz, 2H), 7.05 (d, J = 8.3 Hz, 1H), 7.03-6.98 (m, 1H), 6.72 (d, J = 8.8 Hz, 2H), 4.66-4.63 (m, 2H), 3.67 (s, 3H), 3.54-3.50 (m, 1H), 3.28-3.23 (m, 1H), 3.21 (s, 3H), 2.88-2.82 (m, 1H), 2.56-2.52 (m, 1H), 2.47-2.44 (m, 1H), 1.73 (t, J = 19.0 Hz, 3H); LCMS method: Column = Acquity BEH C18 (50 mm × 2.1 mm, 1.7 μm particles); Mobile phase A = 0.1% formic acid aqueous solution; Mobile phase B = 0.1% formic acid in MeCN solution. Gradient profile (time (min) / %B): 0 / 3, 0.4 / 3, 3.2 / 98, 3.8 / 98, 4.2 / 3, 4.5 / 3; Column temperature = 35°C; Flow rate: 0.6 mL / min. LCMS results: Retention time = 1.93 min; Observed ions = 668.05 (M+H); HPLC Purity = 98%; Chiral HPLC Purity = 96.9%.

[0206] Step 3: Preparation of N-((S)-1-((3P)-3-(4-chloro-1-methyl-3-(methylsulfonamide)-1H-indazole-7-yl)-7-(3,3-difluorobutoxy)-4-oxo-3,4-dihydropyrido[2,3-d]pyrimidine-2-yl)-2-(3,5-difluorophenyl)ethyl)-2-((3bS,4aR)-3-(difluoromethyl)-5,5-difluoro-3b,4,4a,5-tetrahydro-1H-cyclopropa[3,4]cyclopenta[1,2-c]pyrazole-1-yl)acetamide

[0207] [ka]

[0208] At 27°C, (S)-N-((6P)-7-(2-(1-amino-2-(3,5-difluorophenyl)ethyl)-7-(3,3-difluorobutoxy)-4-oxopyrid[2,3-d]pyrimidine-3(4H)-yl)-4-chloro-1-methyl-1H-indazole-3-yl)methanesulfonamide (50g, 74.1 mmol) is mixed with DMF (500mL) and 2-((3bS,4aR)-3-(difluoromethyl)-5,5-difluoro-3b,4 4a,5-tetrahydro-1H-cyclopropa[3,4]cyclopenta[1,2-c]pyrazole-1-yl)acetic acid (21.75 g, 82 mmol), followed by N-(3-dimethylaminopropyl)-N'-ethylcarbodiimide hydrochloride ("EDC-HCl", 18.47 g, 96 mmol), 1-hydroxybenzotriazole hydrate ("HOBt hydrate", 13.62 g, 89 mmol), and N-methylmorpholine (65.2 mL, 593 mmol). The reaction mixture was stirred at 27°C for 16 hours. The progress of the reaction was monitored by TLC (SiO2, 50% siRNA / Pet., Rf=0.5). Upon completion, the reaction mixture was diluted with ice water (1 L), the resulting precipitate was collected by filtration, and then dried under vacuum to obtain the crude product (77 g) as an off-white solid. This crude product was blended with two additional batches of crude product obtained by repeating the procedure on the same scale. Together, 227 g of crude product was purified by silica gel chromatography eluting with a 40-50% ethyl acetate solution in PET. The fraction containing the desired product was pooled and concentrated under reduced pressure to obtain the purified product (180 g). This purified product was blended with an additional batch (25 g) of similarly prepared product.A portion of the purified product (150 g) was further purified in batches (30 × 5 g) by reverse-phase chromatography using the following method (column = RediSep 275 g, HP C18 (CV 243 mL, 150 mL / min); mobile phase A = water:MeCN:TFA (950:50:1); mobile phase B = water:MeCN:TFA (50:950:1); gradient profile (time (min) / %B) = 3 / 10, 6 / 20, 9 / 30, 12 / 40, 15 / 50, 18 / 60, 42 / 70 (compound begins to elute), 52 / 80, 57 / 100; flow rate 80 mL / min; column temperature = 26 °C; input amount = 5 g each time). The fraction containing the pure product was pooled and concentrated under reduced pressure to remove the acetonitrile component. The aqueous solution was made basic by adding saturated NaHCO3, and then extracted with siRNA (3 × 500 mL). The combined organic matter was dried over anhydrous Na2SO4 and then filtered. The filtrate was concentrated under reduced pressure to obtain the desired product (102 g) as an off-white solid. This substance was dissolved in siRNA (200 mL), and then the solution was diluted with n-hexane (1 L). The resulting precipitate was stirred at 27°C for 2 hours and then recovered by filtration. The solid was dried under vacuum. The compound was ground using a mortar and pestle, and then trace amounts of solvent residue were removed by holding the fine solid in a 50°C oven for about 2 hours. This grinding and heating process was repeated (approximately 4-5 times) until all trace solvents were removed (analyzed by NMR) to obtain N-((S)-1-((3P)-3-(4-chloro-1-methyl-3-(methylsulfonamide)-1H-indazole-7-yl)-7-(3,3-difluorobutoxy)-4-oxo-3,4-dihydropyrido[2,3-d]pyrimidine-2-yl)-2-(3,5-difluorophenyl)ethyl)-2-((3bS,4aR)-3-(difluoromethyl)-5,5-difluoro-3b,4,4a,5-tetrahydro-1H-cyclopropa[3,4]cyclopenta[1,2-c]pyrazole-1-yl)acetamide as an off-white solid (88.7g, yield = 87%).1H-NMR (DMSO-d6) δ: 9.86 (s, 1H), 9.45 (d, J = 8.3 Hz, 1H), 8.44 (d, J = 8.7 Hz, 1H), 7.77 (d, J = 8.3 Hz, 1H), 7.48 (d, J = 7.4 Hz, 1H), 7.09 (d, J = 8.7 Hz, 2H), 7.07-6.77 (m, 1H), 6.65 (d, J = 6.2 Hz, 2H), 4.70 (d, J = 16.7 Hz, 1H), 4.65 (t, J = 6.3 Hz, 2H), 4.55 (d, J = 16.7Hz, 1H), 4.51-4.45 (m, 1H), 3.50 (s, 3H), 3.42-3.37 (m, 1H), 3.18 (s, 3H), 3.06-3.00 (m, 1H), 2.56-2.52 (m, 2H), 2.47-2.42 (m, 2H), 1.73 (t, J = 19.2 Hz, 3H), 1.38-1.32 (m, 1H), 0.85-0.81 (m, 1H);LCMS method: Column = Acquity BEH C18 (50 mm × 2.1 mm, 1.7 μm particles), mobile phase A = 0.1% formic acid aqueous solution; mobile phase B = 0.1% formic acid in MeCN solution; gradient profile (time (min) / %B) = 0 / 3, 0.4 / 3, 7.5 / 98, 9.5 / 98, 9.6 / 3, 10 / 3; column temperature = 35°C; flow rate = 0.6 mL / min. LCMS results: retention time = 5.05 min; observed ions = 913.97 (M + H); HPLC purity = 99.5%; chiral HPLC purity = 99.5%.

[0209] Naming of Example 3: The compound of Example 3 prepared above is a homochiral substance containing axial chirality. Axial chirality can be described using the O / M nomenclature detailed in the IUPAC Gold Book (doi:10.1351 / goldbook.A00547). However, at present, the number of software tools capable of generating chemical names including P / M nomenclature is limited, and the options for converting chemical names using this nomenclature into molecular structural representations are even more limited. Therefore, for clarity and convenience, some of the names from Example 3 are shown below.

[0210] The names of Example 3, generated by ChemDraw Professional 16 (without P / M nomenclature), are as follows: N-((S)-1-(3-(4-chloro-1-methyl-3-(methylsulfonamide)-1H-indazole-7-yl)-7-(3,3-difluorobutoxy)-4-oxo-3,4-dihydropyrido[2,3-d]pyrimidine-2-yl)-2-(3,5-difluorophenyl)ethyl)-2-((3bS,4aR)-3-(difluoromethyl)-5,5-difluoro-3b,4,4a,5-tetrahydro-1H-cyclopropa[3,4]cyclopenta[1,2-c]pyrazole-1-yl)acetamide

[0211] The chemical names of Example 3, generated by JChem for Excel (including P / M nomenclature), are as follows: N-[(1S)-1-[(3P)-3-(4-chloro-3-methanesulfonamide-1-methyl-1H-indazole-7-yl)-7-(3,3-difluorobutoxy)-4-oxo-3H,4H-pyrido[2,3-d]pyrimidine-2-yl]-2-(3,5-difluorophenyl)ethyl]-2-[(2S,4R)-9-(difluoromethyl)-5,5-difluoro-7,8-diazatricyclo[4.3.0.0 2 , 4 ]nona-1(6),8-dien-7-yl]acetamide

[0212] The chemical name of Example 3 generated by ChemDraw Professional 16 by manually adding the P / M nomenclature is as follows. N-((S)-1-((3P)-3-(4-chloro-1-methyl-3-(methylsulfonamido)-1H-indazol-7-yl)-7-(3,3-difluorobutoxy)-4-oxo-3,4-dihydropyrido[2,3-d]pyrimidin-2-yl)-2-(3,5-difluorophenyl)ethyl)-2-((3bS,4aR)-3-(difluoromethyl)-5,5-difluoro-3b,4,4a,5-tetrahydro-1H-cyclopropa[3,4]cyclopenta[1,2-c]pyrazol-1-yl)acetamide

[0213] Preparation of N-[(6P)-7-{2-[(1S)-1-amino-2-(3,5-difluorophenyl)ethyl]-7-hydroxy-4-oxo-3H,4H-pyrido[2,3-d]pyrimidine-3-yl}-4-chloro-1-methyl-1H-indazole-3-yl]-N-[(4-methoxyphenyl)methyl]methanesulfonamide

[0214] Scheme

[0215]

Chemical Structure

[0216] Step 1: At -25°C, a suspension of (S)-2-((tert-butoxycarbonyl)amino)-3-(3,5-difluorophenyl)propanoic acid (5.49 g, 18.23 mmol) and 2-amino-6-(benzyloxy)nicotinic acid (4.45 g, 18.23 mmol) in acetonitrile (92 mL) (yellow solution) was mixed with pyridine (9.83 mL, 122 mmol), followed by 2,4,6-tripropyl-1,3,5,2,4,6-trioxatriphosfinan 2,4,6-trioxide ("T3P", 45.2 mL, 76 mmol). The reaction mixture (which became a clear solution after the addition of T3P) was stirred at -25°C to 10°C for 4.5 hours, then N-(7-amino-4-chloro-1-methyl-1H-indazole-3-yl)-N-(4-methoxybenzyl)methanesulfonamide (6 g, 15.19 mmol) was added, and the mixture was stirred for 18 hours while warming to room temperature. The reaction mixture was diluted with ethyl acetate, washed with 1N NaOH, then water, then 0.5 M citric acid, then water, then dried over Na2SO4, and concentrated under vacuum. The resulting residue was purified using a hexane solution of 0-60% ethyl acetate in a 15 CV column, and then by silica (330 g RediSep Gold column) retained with 60% siRNA in a 10 CV column. The desired fraction is pooled and concentrated to obtain tert-butyl N-[(1S)-1-[(3P,3P)-7-(benzyloxy)-3-(4-chloro-3-{N-[(4-methoxyphenyl)methyl]methanesulfonamide}-1-methyl-1H-indazole-7-yl)-4-oxo-3H,4H-pyrido[2,3-d]pyrimidine-2-yl]-2-(3,5-difluorophenyl)ethyl]carbamate (major) and tert-butyl N- A mixture of [(1S)-1-[(3M,3M)-7-(benzyloxy)-3-(4-chloro-3-{N-[(4-methoxyphenyl)methyl]methanesulfonamide}-1-methyl-1H-indazole-7-yl)-4-oxo-3H,4H-pyrido[2,3-d]pyrimidine-2-yl]-2-(3,5-difluorophenyl)ethyl]carbamate (minor) yielded a pale yellow solid (8.1 g, 9.14 mmol, yield 60.1%). LC / MS: m / z = 886.25 [M+1]+.

[0217] Step 2: TFA (21.1 mL, 274 mmol) was added to dichloromethane (45.7 mL) of tert-butyl(S)-(1-(7-(benzyloxy)-3-(4-chloro-3-(N-(4-methoxybenzyl)methylsulfonamide)-1-methyl-1H-indazole-7-yl)-4-oxo-3,4-dihydropyrido[2,3-d]pyrimidine-2-yl)-2-(3,5-difluorophenyl)ethyl)carbamate (product from step 1, 8.1 g, 9.14 mmol) in solution. The mixture was stirred at room temperature for 2 hours. The resulting pale yellow solution was concentrated. The residue was taken to ethyl acetate, then washed three times with 1N NaOH, then dried over Na2SO4, and concentrated under vacuum to obtain an oily residue. The residue was purified by silica gel (330 g RediSep Gold column) using a gradient method (solvent A:solvent B 65:35 → 0:100 (2 CV), then 0:100 (9 CV); solvent A = hexane; solvent B = 9:9:2 hexane:ethyl acetate:MeOH). The first eluted isomer (major) was collected and concentrated under vacuum to obtain N-[(6P)-7-{2-[(1S)-1-amino-2-(3,5-difluorophenyl)ethyl]-7-hydroxy-4-oxo-3H,4H-pyrido[2,3-d]pyrimidine-3-yl}-4-chloro-1-methyl-1H-indazole-3-yl]-N-[(4-methoxyphenyl)methyl]methanesulfonamide (4.1 g, 5.89 mmol, yield 64.5%). 1 H NMR (500 MHz, DMSO-d6) δ 7.86 - 7.98 (m, 1 H) 7.15 - 7.37 (m, 4 H) 6.97 - 7.06 (m, 1 H) 6.70 - 6.89 (m, 4 H) 6.40 - 6.48 (m, 1 H) 4.70 - 4.88 (m, 2 H) 3.41 - 3.81 (m, 7 H) 3.20 - 3.28 (m, 1 H) 3.08 - 3.12 (m, 3 H) 2.71 - 2.79 (m, 1 H) 1.69 - 2.00 (m, 2 H). LC / MS: m / z = 696.20 [M+1] + .

[0218] Preparation of N-((S)-1-((3P)-3-(4-chloro-3-(N-(4-methoxybenzyl)methylsulfonamide)-1-methyl-1H-indazole-7-yl)-7-hydroxy-4-oxo-3,4-dihydropyrido[2,3-d]pyrimidine-2-yl)-2-(3,5-difluorophenyl)ethyl)-2-((3bS,4aR)-3-(difluoromethyl)-5,5-difluoro-3b,4,4a,5-tetrahydro-1H-cyclopropa[3,4]cyclopenta[1,2-c]pyrazole-1-yl)acetamide

[0219]

change

[0220] N-[(6P)-7-{2-[(1S)-1-amino-2-(3,5-difluorophenyl)ethyl]-7-hydroxy-4-oxo-3H,4H-pyrido[2,3-d]pyrimidine-3-yl}-4-chloro-1-methyl-1H-indazole-3-yl]-N-[(4-methoxyphenyl)methyl]methanesulfonamide (0.926 g, 1.330 mmol) is mixed with 2-((3bS,4aR)-3-(difluoromethyl)-5,5- Difluoro-3b,4,4a,5-tetrahydro-1H-cyclopropa[3,4]cyclopenta[1,2-c]pyrazole-1-yl)acetic acid (0.351 g, 1.330 mmol), 2-(3H-[1,2,3]triazolo[4,5-b]pyridine-3-yl)-1,1,3,3-tetramethylisouronium hexafluorophosphate (V) ("HATU", 0.531 g, 1.397 mmol), and DIPEA (0.581 mL, 3.33 mmol) were added. The reaction mixture was stirred for 2 hours, then diluted with water and extracted with ethyl acetate. The combined HCl extract was washed with brine, dried over Na2SO4, and concentrated under vacuum. The crude product was purified by silica gel flash chromatography using a hexane solution of 10-100% ethyl acetate to obtain N-((S)-1-((3P)-3-(4-chloro-3-(N-(4-methoxybenzyl)methylsulfonamide)-1-methyl-1H-indazole-7-yl)-7-hydroxy-4-oxo-3,4-dihydropyrido[2,3-d]pyrimidine-2-yl)-2-(3,5-difluorophenyl)ethyl)-2-((3bS,4aR)-3-(difluoromethyl)-5,5-difluoro-3b,4,4a,5-tetrahydro-1H-cyclopropa[3,4]cyclopenta[1,2-c]pyrazole-1-yl)acetamide (1.1 g, 88%) as an off-white foamy solid. LC / MS: m / z = 942.25 [M+1] + .

[0221] Example 4: Preparation of N-((S)-1-((3P)-3-(4-chloro-1-methyl-3-(methylsulfonamide)-1H-indazole-7-yl)-4-oxo-7)-(3,3,3-trifluoropropoxy)-3,4-dihydropyrido[2,3-d]pyrimidine-2-yl)-2-(3,5-difluorophenyl)ethyl)-2-((3bS,4aR)-3-(difluoromethyl)-5,5-difluoro-3b,4,4a,5-tetrahydro-1H-cyclopropa[3,4]cyclopenta[1,2-c]pyrazole-1-yl)acetamide

[0222] [ka]

[0223] A solution of diisopropyl(E)-diazene-1,2-dicarboxylate ("DIAD", 0.125 mL, 0.637 mmol) in THF (0.2 mL) is prepared by adding N-(1-((3P)-3)-(4-chloro-3-(N-(4-methoxybenzyl)methylsulfonamide)-1-methyl-1H-indazole-7-yl)-7-hydroxy-4-oxo-3,4-dihydropyrido[2,3-d]pyrimidine-2-yl)-2-(3,5-difluorophenyl)ethyl)-2-( (3bS,4aR)-3-(difluoromethyl)-5,5-difluoro-3b,4,4a,5-tetrahydro-1H-cyclopropa[3,4]cyclopenta[1,2-c]pyrazole-1-yl)acetamide (0.2 g, 0.212 mmol), 3,3,3-trifluoropropan-1-ol (0.073 g, 0.637 mmol), and triphenylphosphine (0.178 g, 0.679 mmol) were added dropwise to a mixture of tetrahydrofuran (2.1 mL) at room temperature. The reaction mixture was stirred at room temperature for 18 hours and then concentrated under vacuum. The residue was purified using a hexane gradient of 0-60% ethyl acetate in 15 CV, and then by silica gel (24 g RediSep Gold column) held in a hexane solution of 60% ethyl acetate in 5 CV. The fraction containing the pure product was pooled and concentrated to obtain a yellow solid. The solid was transferred to a solution of DCM (1 mL):TFA (0.5 mL), the solution was cooled to 0°C, and trifluoromethanesulfonic acid (0.057 mL, 0.637 mmol) was added to the solution. The mixture was stirred for 1 hour and then concentrated under vacuum. The residue was transferred to ethyl acetate, washed with 1N NaOH, washed with 0.5 M citric acid, dried over Na2SO4, filtered, and concentrated under vacuum. The residue was subjected to silica gel chromatography (24 g RediSep Gold column) using a hexane solution of 0-60% ethyl acetate in 20 CV, and then 60% ethyl acetate in 10 CV.The fraction containing the pure product was pooled and then concentrated under vacuum to obtain N-(1-((6P)-3-(4-chloro-1-methyl-3-(methylsulfonamide)-1H-indazole-7-yl)-4-oxo-7-(3,3,3-trifluoropropoxy)-3,4-dihydropyrido[2,3-d]pyrimidine-2-yl)-2-(3,5-difluorophenyl)ethyl)-2-((3bS,4aR)-3-(difluoromethyl)-5,5-difluoro-3b,4,4a,5-tetrahydro-1H-cyclopropa[3,4]cyclopenta[1,2-c]pyrazole-1-yl)acetamide (0.078 g, 0.081 mmol, yield 38.0%) as a brown solid. 1 H NMR (500 MHz, METHANOL-d4) δ ppm 8.46 - 8.53 (m, 1 H) 7.28 - 7.34 (m, 1 H) 7.19 - 7.24 (m, 1 H) 7.03 - 7.09 (m, 1 H) 6.53 - 6.81 (m, 4 H) 4.80 (dd, J=5.96, 2.98 Hz, 3 H) 4.49 - 4.62 (m, 2 H) 3.58 - 3.62 (m, 3 H) 3.40 - 3.49 (m, 1 H) 3.22 - 3.24 (m, 3 H) 3.06 - 3.14 (m, 1 H) 2.80 - 2.89 (m, 2H) 2.37 - 2.44 (m, 2 H) 1.32 - 1.37 (m, 1 H) 0.96 - 1.01 (m, 1 H). LCMS analysis method: Column = Acquity UPLC BEH C18, 2.1 × 100 mm, 1.7 μm particles; Injection volume = 5.00 μL; Flow rate = 0.80 mL / min; Solvent A = 95:5 Water:MeCN (containing 0.1% v / v formic acid); Solvent B = 5:95 Water:MeCN (containing 0.1% v / v formic acid); Elution profile = Start %B: 0, End %B: 100, Gradient time: 3.5 min, then hold at 100% B for 1 min; Detection wavelength 1 = 220 nm, Detection wavelength 2 = 254 nm. LCMS retention time = 3.097 min; m / z = 918.05 [M+1] + .

[0224] The compound of Example 3 prepared above is a homochiral substance containing axial chirality. Axial chirality can be described using the O / M nomenclature detailed in the IUPAC Gold Book (doi:10.1351 / goldbook.A00547). However, at present, the number of software tools capable of generating chemical names including P / M nomenclature is limited, and the options for converting chemical names using this nomenclature into molecular structural representations are even more limited. Therefore, for clarity and convenience, some of the names of Example 3 are shown below.

[0225] The names of Example 3, generated by ChemDraw Professional 16 (without P / M nomenclature), are as follows: N-((S)-1-(-3-(4-chloro-1-methyl-3-(methylsulfonamide)-1H-indazole-7-yl)-4-oxo-7)-(3,3,3-trifluoropropoxy)-3,4-dihydropyrido[2,3-d]pyrimidine-2-yl)-2-(3,5-difluorophenyl)ethyl)-2-((3bS,4aR)-3-(difluoromethyl)-5,5-difluoro-3b,4,4a,5-tetrahydro-1H-cyclopropa[3,4]cyclopenta[1,2-c]pyrazole-1-yl)acetamide

[0226] The chemical names of Example 3, generated by JChem for Excel (including P / M nomenclature), are as follows: N-[(1S)-1-[(3P,3P)-3-(4-chloro-3-methanesulfonamide-1-methyl-1H-indazole-7-yl)-4-oxo-7-(3,3,3-trifluoropropoxy)-3H,4H-pyrido[2,3-d]pyrimidine-2-yl]-2-(3,5-difluorophenyl)ethyl]-2-[(2S,4R)-9-(difluoromethyl)-5,5-difluoro-7,8-diazatricyclo[4.3.0.0 2 , 4 ]nona-1(6),8-dien-7-yl]acetamide

[0227] The chemical names for Example 3, generated by ChemDraw Professional 16 with manual addition of P / M nomenclature, are as follows: N-((S)-1-((3P)-3-(4-chloro-1-methyl-3-(methylsulfonamide)-1H-indazole-7-yl)-4-oxo-7)-(3,3,3-trifluoropropoxy)-3,4-dihydropyrido[2,3-d]pyrimidine-2-yl)-2-(3,5-difluorophenyl)ethyl)-2-((3bS,4aR)-3-(difluoromethyl)-5,5-difluoro-3b,4,4a,5-tetrahydro-1H-cyclopropa[3,4]cyclopenta[1,2-c]pyrazole-1-yl)acetamide

[0228] biological methods HIV cell culture assay MT-2 cells, 293T cells and NL 4-3 Proviral DNA clones of the virus were obtained from the NIH AIDS Research and Reference Reagent Program. MT-2 cells were grown in RPMI1640 medium supplemented with 10% heat-inactivated fetal bovine serum (FBS), 100 mg / mL penicillin G, and up to 100 units / mL streptomycin. 293T cells were grown in DMEM medium supplemented with 10% heat-inactivated FBS, 100 mg / mL penicillin G, and up to 100 mg / mL streptomycin. Recombinant NL cells were prepared by replacing a portion of the nef gene with the sea urchin luciferase gene. 4-3 Reference viruses used in these studies were generated using proviral clones. Recombination NL into 293T cells using Transit-293 transfection reagent from Mirus Bio LLC (Madison, WI) 4-3Recombinant viruses were prepared by transfection with proviral clones. After 2-3 days, the supernatant was collected, and the amount of virus present was titrated in MT-2 cells by measuring luciferase enzyme activity using luciferase enzyme activity as a marker. Luciferase was quantified using EnduRen Live Cell Substrate from Promega (Madison, WI). The antiviral activity of the compounds against recombinant viruses was quantified by measuring luciferase activity in MT-2 cells infected with recombinant viruses for 4-5 days in the presence of serially diluted compounds.

[0229] (Fa) = 1 / [1 + (ED 50 By using the exponential form of the median effect equation, which is [ / drug concentration)m], the 50% effective concentration (EC) can be calculated. 50 The following was calculated (Johnson VA, Byington RT. Infectivity Assay. In Techniques in HIV Research. ed. Aldovini A, Walker BD. 71-76. New York: Stockton Press. 1990). Inhibition percentage = 1 / [1+(EC 50 By using the exponential form of the median effect equation, which is [ / drug concentration)m] (where m is a parameter that reflects the slope of the concentration response curve), the 50% inhibitory concentration (EC2) can be calculated. 50 ) was calculated.

[0230] Compound cytotoxicity and corresponding CC 50 The values ​​were determined using the same protocol as described in the antiviral assay, except that uninfected cells were used. Cytotoxicity was assessed on day 4 in uninfected MT-2 cells using a colorimetric assay based on XTT (2,3-bis[2-methoxy-4-nitro-5-sulfophenyl]-2H-tetrazolium-5-carboxyanilide intramolecular salt) (Sigma-Aldrich, St Louis, Mo).

[0231] [Table 1]

[0232] Procedure for measuring pharmacokinetic parameters in subcutaneous in vivo experiments (Formulation A) PEG300 (0.906 mL) was added to a 20 mL vial containing Example 1 (320 mg). The mixture was sonicated to obtain a clear solution. Water (0.160 mL) was added to this solution to obtain a slightly cloudy solution. The mixture was sonicated for 5 minutes to obtain a clear solution. The resulting solution is "Formulation A," and its concentrations are 21.3 w / w% of Example 1, 68.1 w / w% of PEG300, and 10.6 w / w% of water.

[0233] "Formulation A" was administered to Wisterhan rats by subcutaneous injection at a dose of 0.167 mL / kg. Blood samples were collected at 0.5 hours, 1 hour, 3 hours, 5 hours, 7 hours, 24 hours, 48 ​​hours, 72 hours, and on days 6, 8, 12, 15, 19, 22, 26, 29, 33, 36, 40, 43, 47, 50, 54, 57, and 61 after administration. Blood samples were collected in K2EDTA tubes and centrifuged at 1500-2000 × g to obtain plasma. Plasma samples were stored at -80°C until analysis by LC-MS / MS. All in vitro samples were injected into an MDS Sciex API 4000 triple quadrupole LC-MS / MS system. The analytical column used was a Phenomenex Kinetex® 2.6 μm PS (C18, 2.1 mm × 50 mm, 2.6 μm) maintained at room temperature. Mobile phase A consisted of a 0.1% (v / v) formic acid MilliQ purified aqueous solution. Mobile phase B consisted of a 0.1% (v / v) formic acid acetonitrile solution. The flow rate was 0.70 mL / min. The gradient was maintained at 35% for 0.5 minutes for mobile phase B, then linearly increased from 35% to 98% over 1.5 minutes, maintained at 98% for 0.5 minutes, and maintained at 35% for 0.5 minutes. The results of the PK experiment are shown in Table 1 and Figure 1.

[0234] [Table 2]

[0235] This disclosure is not limited to the exemplary embodiments described above, and the embodiments should be considered illustrative rather than restrictive in all respects. References are made to the appended claims rather than to the embodiments described above, and therefore all modifications that fall within the meaning and scope of equivalence to the claims are intended to be encompassed.

Claims

1. Formula Ia, Formula Ib, Formula Ic, or Formula Id: 【Chemistry 1】 A pharmaceutical composition comprising a compound or a pharmaceutically acceptable salt thereof, and a solvent or diluent selected from the group consisting of water, alcohol, polyethylene glycol (PEG), N-methyl-2-pyrrolidone (NMP), ethyl lactate, propylene glycol, glycoflor, and dimethyl sulfoxide (DMSO).

2. The pharmaceutical composition according to claim 1, further comprising an additive selected from the group consisting of polysorbate 20, polysorbate 80, poloxamer 188, poloxamer 207, poloxamer 338, sodium chloride, and sodium hydroxide.

3. A pharmaceutical composition according to claim 1 or 2, which is a solution.

4. A pharmaceutical composition according to claim 1 or 2, which is a suspension.

5. A pharmaceutical composition according to any one of claims 1 to 4, comprising water.

6. A pharmaceutical composition according to any one of claims 1 to 5, comprising polyethylene glycol (PEG).

7. The pharmaceutical composition according to claim 6, wherein the average molecular weight of PEG is approximately 300 (PEG300).

8. A pharmaceutical composition according to any one of claims 1 to 7, comprising ethanol.

9. A pharmaceutical composition according to any one of claims 2 to 8, comprising poloxamer 188.

10. A pharmaceutical composition according to any one of claims 2 to 9, comprising sodium hydroxide.

11. The pharmaceutical composition according to any one of claims 2 to 10, wherein at least 90% by weight of the solvent or diluent is water and PEG300.

12. The pharmaceutical composition according to any one of claims 2 to 11, wherein at least 90% by weight of the additive is poloxamer 188.

13. The pharmaceutical composition according to claim 11, wherein the water is present in the pharmaceutical composition in an amount of 8 to 20% by weight, and the PEG 300 is present in the pharmaceutical composition in an amount of 60 to 85% by weight.

14. The pharmaceutical composition according to claim 13, wherein the water is present in the pharmaceutical composition in an amount of 8 to 12% by weight, and the PEG 300 is present in the pharmaceutical composition in an amount of 63 to 70% by weight.

15. The pharmaceutical composition according to any one of claims 1 to 14, wherein a compound of formula Ia, formula Ib, formula Ic, or formula Id, or a pharmaceutically acceptable salt thereof, is present in the pharmaceutical composition at a concentration of 50 to 500 mg / mL by weight of the free compound of formula Ia, formula Ib, formula Ic, or formula Id.

16. The pharmaceutical composition according to claim 15, wherein a compound of formula Ia, formula Ib, formula Ic, or formula Id, or a pharmaceutically acceptable salt thereof, is present in the pharmaceutical composition at a concentration of 225 to 275 mg / mL by weight, based on the free compound of formula Ia, formula Ib, formula Ic, or formula Id.

17. The pharmaceutical composition according to claim 15, wherein a compound of formula Ia, formula Ib, formula Ic, or formula Id, or a pharmaceutically acceptable salt thereof, is present in the pharmaceutical composition at a concentration of 275 to 300 mg / mL by weight, based on the free compound of formula Ia, formula Ib, formula Ic, or formula Id.

18. The pharmaceutical composition according to claim 15, wherein a compound of formula Ia, formula Ib, formula Ic, or formula Id, or a pharmaceutically acceptable salt thereof, is present in the pharmaceutical composition at a concentration of 350 to 425 mg / mL by weight, based on the free compound of formula Ia, formula Ib, formula Ic, or formula Id.

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