A plasmin inhibitor, its preparation method and application

Abstract

The present invention relates to a plasmin inhibitor, a process for its preparation and use in the pharmaceutical field. The present invention also provides a process for preparing the compounds of the invention, compositions comprising the compounds of the invention and use as plasmin inhibitors and pharmaceutical uses thereof.

Description

Technical Field

[0001] This invention relates to the field of pharmaceutical chemistry, specifically to a plasmin inhibitor, its preparation method, and its application in the pharmaceutical field. Background Technology

[0002] Plasmin is a proteolytic enzyme that degrades fibrin. When tissue damage causes blood vessel rupture, it triggers hemostasis mechanisms: vasoconstriction, platelet embolism, and the initiation of coagulation, ultimately leading to the formation of stable fibrin. Simultaneously, due to fibrin deposition, the fibrinolytic system is activated. This system maintains a balance between fibrin formation and breakdown, playing a role in maintaining vascular patency and remodeling damaged tissue during the repair of damaged blood vessel walls (Tengborn L, M, Berntorp E. Thromb Res. 2015 Feb; 135(2):231-42).

[0003] The fibrinolytic system includes plasminogen, tissue-type plasminogen activator (tPA), and urokinase-type plasminogen activator (uPA). Plasminogen binds to lysine residues on the fibrin surface and is converted into plasmin via an activator (i.e., tPA) released from endothelial cells. Fibrinolysis inhibition can be used to treat bleeding. The use of antifibrinolytic drugs can reduce blood loss in cardiac surgery, trauma, orthopedic surgery, solid organ transplantation, obstetrics and gynecology, neurosurgery, and non-surgical conditions (Ng W, Jerath A, ...). M. Anaesthesiol Intensive Ther. 2015; 47(4):339-50). In the early 1950s, it was discovered that lysine amino acids inhibited the activation of plasminogen, but the effect was too weak to be used to treat fibrinolytic hemorrhage. In 1953, Shosuke Okamoto et al. showed that several thiol groups and aminocarbonates had anti-plasma protein effects, and found that the synthetic derivative of lysine, ε-aminohexanoic acid (EACA), had a strong inhibitory effect on plasminogen. EACA has been widely used in clinical practice, but in addition to mild gastrointestinal side effects such as nausea, a larger dose is required. In 1962, 4-amino-methyl-cyclohexane-carbonic acid (AMCHA) was discovered. This compound contains two stereoisomers. Further studies showed that its trans form (trans-4-aminomethylcyclohexanecarboxylic acid, i.e., tranexamic acid, TXA) has anti-fibrinolytic ability, with an activity about 10 times that of EACA, and has been shown to have stronger tolerance (Tengborn L, M, Berntorp E. Thromb Res. 2015 Feb; 135(2):231-42).

[0004] Tranexamic acid is a synthetic lysine derivative and antifibrinolytic agent that forms a reversible complex with plasminogen. By binding to plasminogen, it blocks the interaction between plasminogen and the plasmin heavy chain and fibrin lysine residues, thereby preventing plasminogen from binding to the fibrin surface and thus delaying fibrinolysis. Tranexamic acid has been approved for the treatment of severe menstrual bleeding and various surgical bleeding disorders, and is currently the most commonly used hemostatic drug in clinical practice. However, numerous literature reports indicate that oral administration of tranexamic acid easily causes gastrointestinal adverse reactions such as nausea, vomiting, diarrhea, and indigestion, and its dosage is relatively high; patients may also experience complications such as epilepsy after taking the drug.

[0005] Other similar hemostatic drugs, such as aminocaproic acid, have problems such as rapid excretion from the body, weak hemostatic effect, short duration of action, and more toxic reactions. Excessive dosage can lead to thrombosis, limiting their use in patients with a tendency to thrombosis, a history of thrombotic vascular disease, or renal insufficiency. Tranexamic acid has a similar mechanism to aminocaproic acid but is 4-5 times stronger. It is effective for general chronic bleeding but has no hemostatic effect on traumatic bleeding or cancerous bleeding. Furthermore, excessive dosage can also promote thrombosis. Aprotinin, a commonly used hemostatic drug in coronary artery bypass surgery, was withdrawn from the market by the FDA in 2008 due to its potential to induce renal failure, myocardial infarction, and heart failure.

[0006] Other hemostatic drugs with different mechanisms of action, such as carbazoline which acts on blood vessels, can induce epilepsy with repeated use; thrombin, a hemostatic drug that promotes the coagulation process, can only be used for gastrointestinal bleeding or local bleeding.

[0007] Given the very limited selection of hemostatic drugs available in clinical practice, and the deficiencies in dosage and clinical indications, as well as the problems of high dosage, numerous adverse reactions, and the potential to cause complications such as epilepsy, it is necessary to develop a new hemostatic drug to better meet clinical needs. Summary of the Invention

[0008] One of the objectives of this invention is to overcome the aforementioned deficiencies of the prior art and to provide a novel compound with coagulation and hemostatic activities.

[0009] Specifically, the present invention provides compounds of Formula I, including pharmaceutically acceptable salts, hydrates, isomers, prodrugs, and mixtures thereof:

[0010]

[0011] Wherein, X is selected from CH2, NR3, O, and S; Y is selected from CH and N; in some specific implementation schemes, X is NR3 and Y is N;

[0012] R1 is selected from hydrogen, NRaRb, hydroxyl, substituted or unsubstituted aryl, substituted or unsubstituted alkyl, substituted or unsubstituted aromatic heterocyclic groups; wherein Ra and Rb are each independently selected from hydrogen, alkyl, or haloalkyl.

[0013] R2 is selected from hydrogen, substituted or unsubstituted alkyl, haloalkyl, substituted or unsubstituted cycloalkyl, substituted or unsubstituted alicyclic, substituted or unsubstituted aryl, substituted or unsubstituted aromatic heterocyclic;

[0014] R3 is selected from hydrogen, substituted or unsubstituted alkyl, substituted or unsubstituted cycloalkyl, substituted or unsubstituted alicyclic, substituted or unsubstituted aryl, substituted or unsubstituted aromatic heterocyclic, -(CH2)mNRcRd, where m = 1, 2, 3, 4, and Rc and Rd are each independently selected from hydrogen, alkyl, or haloalkyl.

[0015] In some specific embodiments, R1 in this invention is selected from hydrogen, NRaRb, hydroxyl, substituted or unsubstituted 6-10 aryl, substituted or unsubstituted C1-C4 alkyl, substituted or unsubstituted 6-10 aryl heterocyclic group; wherein Ra and Rb are each independently selected from hydrogen, alkyl, haloalkyl;

[0016] In some specific embodiments, R2 of the present invention is selected from hydrogen, substituted or unsubstituted C1-C4 alkyl, C1-C4 haloalkyl, substituted or unsubstituted C3-C6 cycloalkyl, substituted or unsubstituted 4-8 membered alicyclic group, substituted or unsubstituted 6-10 membered aryl group, and substituted or unsubstituted 6-10 membered aromatic heterocyclic group.

[0017] In some specific embodiments, R3 of the present invention is selected from hydrogen, substituted or unsubstituted C1-C4 alkyl, C1-C4 haloalkyl, substituted or unsubstituted C3-C6 cycloalkyl, substituted or unsubstituted 4-8 membered alicyclic group, substituted or unsubstituted 6-10 membered aryl group, and substituted or unsubstituted 6-10 membered aromatic heterocyclic group.

[0018] In some specific embodiments, the alkyl group of the present invention is selected from methyl, ethyl, propyl, isopropyl, n-butyl, and tert-butyl; the alkyl group may be selectively substituted by one or more groups selected from halogen, amino, hydroxyl, cycloalkyl, and alicyclic groups;

[0019] In some specific embodiments, the cycloalkyl group of the present invention is selected from cyclopropyl, cyclobutyl, cyclopentyl, cyclopentenyl, cyclohexyl, and cyclohexenyl.

[0020] In some specific embodiments, the alicyclic group of the present invention is selected from oxadiazonyl, pyrrolidinyl, tetrahydrofuranyl, tetrahydropyranyl, tetrahydrothiophenyl, piperidinyl, and morpholinyl.

[0021] In some specific embodiments, the aryl group of the present invention is selected from phenyl and naphthyl groups;

[0022] In some specific embodiments, the aromatic heterocyclic group of the present invention is selected from pyridinyl, pyrimidinyl, pyridazinyl, imidazoleyl, pyrazolyl, thiazolyl, thiophenyl, oxazolyl, isoxazolyl, and 1,2,4-oxadiazolyl.

[0023] In some specific embodiments, R1 is selected from hydroxyl groups, and R2 is selected from hydrogen.

[0024] In some specific embodiments, X in this invention is selected from CH2, NR3, O, and S, wherein R3 is selected from hydrogen, -Q-(R Q ) n Q is selected from C1-C4 alkyl, C3-C6 cycloalkyl, and 4-6 membered saturated alicyclic alkyl groups; n is 0 or 1; R Q The group is selected from methyl, phenyl, tetrahydropyranyl, morpholinyl, piperidinyl, methylpiperidinyl, o-chlorophenyl, carboxyl, hydroxyl, dimethylamino, and halogen; further, Q is preferably selected from methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, tert-butyl, cyclopropane, cyclobutane, cyclopentane, piperidinyl, tetrahydropyranyl, morpholinyl, and cyclopropoxy.

[0025] In some specific embodiments, X in this invention is preferably selected from CH2, NR3, O, and S, wherein R3 is selected from hydrogen, tetrahydropyranyl, tetrahydropyranmethyl, methyl, ethyl, n-propyl, isopropyl, tert-butyl, trifluoromethyl, trifluoroethyl, benzyl, phenethyl, phenyl, piperidinylmethyl, methylpiperidinylmethyl, hydroxyethyl, hydroxymethyl, carboxymethyl, carboxyethyl, piperidinyl, methylpiperidinyl, piperidinylmethyl, cyclopropyl, cyclopentyl, cyclobutyl, dimethylaminopropyl, dimethylaminoethyl, glycidyl, and o-chlorophenylethyl.

[0026] More preferably from CH2, NR3, O, S, wherein R3 is selected from hydrogen, methyl, ethyl, isopropyl, tert-butyl, tetrahydropyranyl, tetrahydropyranmethyl, trifluoromethyl, trifluoroethyl, benzyl, phenethyl, phenyl, piperidinemethyl, methylpiperidinemethyl, hydroxyethyl, hydroxymethyl, carboxymethyl, carboxyethyl, piperidine;

[0027] Further preferred from CH2, NR3, O, S, wherein R3 is selected from hydrogen, tetrahydropyranyl, tetrahydropyranmethyl, methyl, ethyl, tert-butyl, and carboxymethyl;

[0028] It is particularly preferred to be selected from O, NR3, wherein the R3 is selected from hydrogen, tetrahydropyranyl, and tetrahydropyranmethyl.

[0029] In some specific embodiments, Y in this invention is preferably selected from N;

[0030] In some embodiments, the tetrahydropyranmethyl group of the present invention may be tetrahydropyran-4-ylmethyl, tetrahydropyran-2-ylmethyl, or tetrahydropyran-3-ylmethyl; in some specific embodiments, the tetrahydropyranmethyl group of the present invention is preferably tetrahydropyran-4-ylmethyl.

[0031] In some embodiments, the tetrahydropyranyl group of the present invention may be tetrahydropyran-4-yl, tetrahydropyran-2-yl, or tetrahydropyran-3-yl; in some specific embodiments, the tetrahydropyranyl group of the present invention is preferably tetrahydropyran-4-yl.

[0032] In some embodiments, the piperidinyl group of the present invention may be piperidin-4-yl, piperidin-2-yl, or piperidin-3-yl; in some specific embodiments, the piperidinyl group of the present invention is preferably piperidin-4-yl.

[0033] In some embodiments, the piperidine methyl group of the present invention may be piperidine-4-ylmethyl, piperidine-2-ylmethyl, or piperidine-3-ylmethyl; in some specific embodiments, the piperidine methyl group of the present invention is preferably piperidine-4-ylmethyl.

[0034] In some embodiments, the methylpiperidinemethyl or methylpiperidinyl group of the present invention refers to the substitution of methyl at any position on the piperidine ring of the piperidinemethyl or piperidinyl group; in some specific embodiments, the methylpiperidinemethyl or preferably N-methylpiperidinemethyl group of the present invention, wherein the methylpiperidinyl group is preferably N-methylpiperidinyl group;

[0035] In some embodiments, the phenethyl group of the present invention may be 2-phenylethyl or 1-phenylethyl; in some specific embodiments, the phenethyl group of the present invention is preferably 1-phenylethyl.

[0036] In some embodiments, the trifluoroethyl group of the present invention may be 2,2,2-trifluoroethyl, 1,2,2-trifluoroethyl, or 1,1,2-trifluoroethyl; in some specific embodiments, the trifluoroethyl group of the present invention is preferably 2,2,2-trifluoroethyl.

[0037] In some specific embodiments, the compound of the present invention has the following structure:

[0038]

[0039]

[0040] Another object of the present invention is to provide a pharmaceutical composition comprising at least one of the aforementioned compounds, or a pharmaceutically acceptable salt, hydrate, isomer, prodrug, or mixture thereof, and at least one pharmaceutically acceptable excipient.

[0041] Another object of the present invention is to provide the aforementioned compound or its pharmaceutically acceptable salts, hydrates, isomers, prodrugs, mixtures, or pharmaceutical compositions for use in the preparation of a medicament. The medicament possesses therapeutic activities of coagulation and hemostasis, and can be used for abnormal bleeding caused by hyperfibrinolysis, surgical and postoperative bleeding, etc.

[0042] Another object of the present invention is to provide a method for treating and / or alleviating bleeding disorders or conditions, comprising administering to a patient the foregoing one or more of the aforementioned pharmaceutical compositions or compounds of Formula I or thereof, or pharmaceutically acceptable salts, hydrates, isomers, prodrugs or mixtures thereof.

[0043] Terminology Definition

[0044] Unless otherwise stated, the following terms and phrases as used herein are intended to have the following meanings. A particular term or phrase should not be considered uncertain or unclear unless specifically defined, but should be understood in its ordinary sense. When a trade name appears herein, it is intended to refer to the corresponding product or its active ingredient.

[0045] The term "pharmaceutically acceptable" as used here means that it is intended for use only in contact with human and animal tissues without excessive toxicity, irritation, allergic reactions or other problems or complications, in proportion to a reasonable benefit / risk ratio.

[0046] The term "pharmaceutically acceptable salt" refers to the salt of the compounds of this invention, prepared by reacting a compound with a relatively non-toxic acid or base, as discovered in this invention, with a specific substituent. When the compounds of this invention contain relatively acidic functional groups, base addition salts can be obtained by contacting the neutral form of such compounds with a sufficient amount of base in a pure solution or a suitable inert solvent. When the compounds of this invention contain relatively basic functional groups, acid addition salts can be obtained by contacting the neutral form of such compounds with a sufficient amount of acid in a pure solution or a suitable inert solvent.

[0047] The compounds of the present invention may exist in specific geometric or stereoisomeric forms. The present invention envisions all such compounds, including cis and trans isomers, (-)- and (+)- enantiomers, (R)- and (S)- enantiomers, diastereomers, (D)- isomers, (L)- isomers, racemic mixtures thereof, and other mixtures, all of which are within the scope of the present invention.

[0048] "Alkyl" refers to a straight-chain or branched saturated aliphatic hydrocarbon group. For example, C1-C4 alkyl refers to a saturated aliphatic hydrocarbon group containing 1 to 4 carbon atoms, including but not limited to methyl, ethyl, propyl, isopropyl, n-butyl, tert-butyl, and their various isomers.

[0049] "Cycloalkyl" refers to a monocyclic or polycyclic cyclic hydrocarbon substituent that is saturated or partially unsaturated. For example, "C3-C6 cycloalkyl" refers to a cycloalkyl group containing 3 to 6 carbon atoms. Typical C3-C6 cycloalkyl groups include, but are not limited to, cyclopropyl, cyclobutyl, cyclopentyl, cyclopentenyl, cyclohexyl, and cyclohexenyl.

[0050] "Aliphatic heterocyclic group" refers to a saturated monocyclic hydrocarbon substituent in which one or more ring atoms are replaced by heteroatoms selected from N, O, and S, and the remaining ring atoms are carbon. For example, "3-8 membered aliphatic heterocycle" refers to a saturated cyclic hydrocarbon substituent containing 3-8 ring atoms, in which one or more ring atoms are replaced by heteroatoms selected from N, O, and S, and the remaining ring atoms are carbon. Specific examples include, but are not limited to: oxabutyl, pyrrolidinyl, tetrahydrofuranyl, morpholinyl, etc.

[0051] "Aromatic heterocyclic group" refers to an aromatic cyclic substituent in which one or more ring atoms are replaced by a heteroatom selected from N, O, and S, and the remaining ring atoms are carbon. For example, "5-6 membered aromatic heterocyclic group" refers to an aromatic heterocyclic group containing 5 to 6 ring atoms, and specific examples include, but are not limited to, pyridinyl, pyrimidinyl, pyridazinyl, imidazoleyl, pyrazolyl, thiazolyl, oxazolyl, isoxazolyl, and 1,2,4-oxadiazolyl.

[0052] "Alicyclic substituent" refers to a saturated cyclic substituent in which one or more ring atoms are replaced by heteroatoms selected from N, O, and S. For example, "3-5 membered heterocyclic substituent" refers to a saturated or unsaturated cyclic substituent containing 3 to 5 ring atoms, in which one or more ring atoms are replaced by heteroatoms selected from N, O, and S. Specific examples include, but are not limited to: oxocyclic butyl, azircyclic butyl, tetrahydrofuranyl, pyrrolidinyl, thiazolyl, etc.

[0053] "Aryl" refers to an aromatic cyclic group. For example, "6-10 aryl" refers to an aromatic cyclic group containing 6 to 10 carbon ring atoms. Examples of aryl moiety include phenyl, naphthyl, etc.

[0054] "Optional" means that the event or situation described below may occur but is not required to occur.

[0055] All abbreviations used in this invention are known to those skilled in the art and, unless otherwise stated, represent the meanings known in the art. For example: DMF refers to N,N-dimethylformamide; THF refers to tetrahydrofuran; and Me refers to methyl.

[0056] Experiments have shown that the compound of this invention has excellent coagulation and hemostatic activity, which is significantly better than tranexamic acid, the most widely used hemostatic drug in clinical practice, and has great clinical application value.

[0057] The abbreviations used in this invention have the following meanings:

[0058]

[0059] Detailed Implementation

[0060] The following examples illustrate the synthesis methods of the compounds and intermediates of this invention. These examples are merely illustrative and should not be construed as limiting the scope of the invention. Unless otherwise specified, the raw materials and reagents involved in this invention are commercially available, and the specific source does not affect the implementation of the technical solution of this invention.

[0061] Example 1: Preparation of (4-amino-3,4-dihydro-2H-pyrano[2,3-b]pyridin-7-yl)phosphonate

[0062] Step 1: Preparation of (3-((tert-butyldimethylsilyl)oxy)propylidene)-2-methylpropane-2-sulfinamide

[0063] 5.6 g of 3-((tert-butyldimethylsilyl)oxy)propionaldehyde was dissolved in 50 mL of dichloromethane, and 5.4 g of tert-butylsulfinamide and 23.8 g of anhydrous copper sulfate were added. The mixture was stirred at room temperature for 48 h. TLC showed that the starting material was completely consumed. The mixture was filtered through diatomaceous earth and purified by column chromatography to obtain the title compound (7.35 g).

[0064]

[0065] MS(ESI)m / z(M+H) + =292.1.

[0066] Step 2: Preparation of 3-((tert-butyldimethylsilyl)oxy)-1-(2,6-dichloropyridin-3-yl)propyl)-2-methylpropane-2-sulfinamide

[0067] 3.74 g of 2,6-dichloropyridine was weighed into a 250 mL three-necked flask and dissolved in 50 mL of tetrahydrofuran. The mixture was purged three times with argon gas and cooled to -78 °C. Then, 12.5 mL of diisopropylaminolithium (LDA) was slowly added over 10 min. After stirring for 30 min, a solution of N-(3-((tert-butyldimethylsilyl)oxy)propylidene)-2-methylpropane-2-sulfinamide (7.35 g) dissolved in 10 mL of tetrahydrofuran was slowly added dropwise using a syringe. The reaction was carried out at -78 °C for 2 h. TLC showed that the starting material was completely consumed. The mixture was quenched with water, extracted twice with ethyl acetate, dried over anhydrous sodium sulfate, and purified by column chromatography to obtain the title compound (2.6 g).

[0068]

[0069] MS(ESI)m / z(M+H) + =438.9.

[0070] Step 3: Preparation of 1-(2,6-dichloropyridin-3-yl)-3-hydroxypropyl)-2-methylpropane-2-sulfinamide

[0071] 2.6 g of 3-((tert-butyldimethylsilyl)oxy)-1-(2,6-dichloropyridin-3-yl)propyl)-2-methylpropane-2-sulfinamide was weighed and dissolved in 30 mL of tetrahydrofuran and cooled to 0 °C. 17.8 mL of 1 M tetrabutylammonium fluoride solution was added, and the reaction was carried out at 0 °C for 1 h. TLC showed that the starting material was completely consumed. The solution was quenched with water, extracted twice with ethyl acetate, washed twice with saturated brine, dried over anhydrous sodium sulfate, and purified by column chromatography to obtain the title compound (1.6 g).

[0072]

[0073] MS(ESI)m / z(M+H) + =325.0.

[0074] Step 4: Preparation of 7-chloro-3,4-dihydro-2H-pyrano[2,3-b]pyridin-4-yl)-2-methylpropane-2-sulfinamide

[0075] 1-(2,6-dichloropyridin-3-yl)-3-hydroxypropyl)-2-methylpropane-2-sulfinamide (200 mg) and potassium tert-butoxide (173 mg) were weighed into tert-butanol (5 mL) and reacted at 85 °C for 2 h. TLC showed that the starting material was completely consumed. The reaction was quenched with water, extracted twice with dichloromethane, dried over anhydrous sodium sulfate, and purified by column chromatography to obtain the title compound (133 mg).

[0076]

[0077] MS(ESI)m / z(M+H) + =289.1.

[0078] Step 5: Preparation of diethyl 4-((-tert-butylsulfinyl)amino)-3,4-dihydro-2H-pyrano[2,3-b]pyridin-7-yl)phosphonate

[0079] 50 mg of -N-(7-chloro-3,4-dihydro-2H-pyrano[2,3-b]pyridin-4-yl)-2-methylpropane-2-sulfinamide was dissolved in toluene (2 mL). Tris(dibenzylacetone)dipalladium (16 mg), 1,1'-bis(diphenylphosphine)ferrocene (19 mg), triethylamine (53 mg), and diethyl phosphite (48 mg) were added sequentially. The mixture was purged three times with argon gas and reacted at 120 °C for 2.5 h. TLC showed that the starting material was completely consumed. The mixture was quenched with water, extracted twice with ethyl acetate, dried over anhydrous sodium sulfate, and purified by Prep-TLC to obtain the title compound (13 mg).

[0080]

[0081] MS(ESI)m / z(M+H) + =391.0.

[0082] Step 6: Preparation of (4-amino-3,4-dihydro-2H-pyrano[2,3-b]pyridin-7-yl)phosphonate

[0083] 13 mg of diethyl (4-((tert-butylsulfinyl)amino)-3,4-dihydro-2H-pyrano[2,3-b]pyridin-7-yl)phosphonate was dissolved in 3 mL of concentrated hydrochloric acid and reacted at 100 °C for 3 h. LCMS showed that the starting material was completely consumed. The mixture was concentrated under reduced pressure and purified by pre-HPLC to obtain the title compound (6.5 mg).

[0084]

[0085] MS(ESI)m / z(M+H) + =231.0.

[0086] 1 H NMR (400MHz, Deuterium Oxide)δ8.34(dd,J=8.0,3.1Hz,1H),7.70(t,J=7.3Hz,1H),4.78(s,1H),4.72(q,J=3. 9,3.2Hz,2H),2.57(ddd,J=13.4,9.5,5.8Hz,1H),2.39(ddt,J=15.7,10.8,4.7Hz,1H).

[0087] Example 2: Preparation of (4-amino-3,4-dihydro-2H-benzopyran-7-yl)phosphonate

[0088]

[0089] Step 1: Preparation of N-(7-bromobenzopyran-4-alkylene)-2-methylpropane-2-sulfinamide

[0090] 7-Bromo-4-dihydrochromone (2.5 g) was dissolved in tetrahydrofuran (4 mL), followed by the addition of tert-butylsulfinamide (2.0 g) and tetraethyl titanate (4 mL). The reaction was carried out overnight at 85 °C, and TLC showed complete reaction of the starting material. The mixture was quenched with water, filtered, and the filter cake was washed with ethyl acetate. The filtrate was backwashed twice with saturated brine (2 × 15 mL). The organic phases were combined, dried over anhydrous sodium sulfate, and purified by column chromatography to give the title compound (2.1 g).

[0091]

[0092] MS(ESI)m / z(M+H) + =329.9.

[0093] Step 2: Preparation of N-(7-bromobenzopyran-4-yl)-2-methylpropane-2-sulfinamide

[0094] 1.4 g of N-(7-bromobenzopyran-4-alkylene)-2-methylpropane-2-sulfinamide was dissolved in 10 mL of tetrahydrofuran. 10.6 mL of 1.0 M trisec-butylborohydride was slowly added at 0 °C. The system was slowly heated to room temperature and reacted for 3 h. TLC showed that the reaction was complete. The reaction was quenched with an aqueous solution of ammonium chloride. The mixture was extracted twice with ethyl acetate, dried over anhydrous sodium sulfate, and purified by column chromatography to give the title compound (738 mg).

[0095]

[0096] MS(ESI)m / z(M+H) + =332.0.

[0097] Step 3: Preparation of diethyl (4-((tert-butylsulfinyl)amino)benzodihydropyran-7-yl)phosphonate

[0098] 0.32 g of N-(7-bromobenzopyran-4-yl)-2-methylpropane-2-sulfinamide was weighed into a microwave reactor. N,N-diisopropylethylamine (0.21 mL), diethyl phosphite (0.16 mL), palladium acetate (8 mg), and dppf (21 mg) were added sequentially. The reactor was purged three times with argon gas. 3 mL of anhydrous N,N-dimethylformamide / ethylene glycol dimethyl ether (9:1, v / v) was added. The reaction was carried out at 115 °C for 25 min in a microwave reactor. TLC showed complete reaction of the starting material. The reaction was quenched with water, extracted twice with ethyl acetate, dried over anhydrous sodium sulfate, and purified by column chromatography to obtain the title compound (233 mg).

[0099]

[0100] MS(ESI)m / z(M+H) + =390.0.

[0101] Step 4: Preparation of (4-amino-3,4-dihydro-2H-benzopyran-7-yl)phosphonate

[0102] Weigh 0.145 g of diethyl (4-((tert-butylsulfinyl)amino)benzodihydropyran-7-yl)phosphonate and dissolve it in 4 mL of 12 M concentrated hydrochloric acid. Heat the system to 100 °C and react overnight. LC-MS showed that the reaction of the starting material was complete. After concentration under reduced pressure, the compound was purified by pre-HPLC to obtain the title compound (39 mg).

[0103]

[0104] MS(ESI)m / z(MH) - =227.9.

[0105] 1 H NMR (400MHz, Deuterium Oxide)δ7.32(dd,J=7.9,4.4Hz,1H),7.20(dd,J=12.5,7.9Hz,1H),7.11(d,J=14.7Hz,1H),4.52(d,J= 5.6Hz,1H),4.17(tdt,J=12.1,9.1,3.6Hz,2H),2.29(ddd,J=14.4,9.5,4.7Hz,1H),2.12-2.01(m,1H).

[0106] Example 3: Preparation of (4-amino-1,2,3,4-tetrahydroquinoline-7-yl)phosphonate

[0107]

[0108] Step 1: Preparation of methyl 3-((3-bromophenyl)amino)propionate

[0109] m-Bromoaniline (9 g), methyl acrylate (4.6 g), and glacial acetic acid (297 μL) were placed in a sealed tube and reacted at 110 °C for 12 h. After the starting material was consumed by TLC, the mixture was diluted with ethyl acetate, washed three times with water, dried over anhydrous sodium sulfate, and concentrated. The crude product was purified by column chromatography to give the title compound (10 g).

[0110]

[0111] MS(ESI)m / z(M+H) + =257.9.

[0112] Step 2: Preparation of methyl 3-(N-(3-bromophenyl)-4-methylphenylsulfonamido)propionate

[0113] Methyl 3-((3-bromophenyl)amino)propionate (10 g) and 4-toluenesulfonyl chloride (8.1 g) were dissolved in pyridine (20 mL) and reacted at room temperature for 2 h. After the starting materials were consumed by TLC, the reaction solution was concentrated to dryness, dissolved and diluted with ethyl acetate, washed once with hydrochloric acid (1 N) and once with saturated brine, and then dried over anhydrous sodium sulfate and concentrated. The crude product was purified by column chromatography to give the title compound (15.6 g).

[0114]

[0115] MS(ESI)m / z(M+H) + =411.9.

[0116] Step 3: Preparation of 3-(N-(3-bromophenyl)-4-methylphenylsulfonamido)propionic acid

[0117] Methyl 3-(N-(3-bromophenyl)-4-methylphenylsulfonamido)propionate (15.6 g) was dissolved in a solution of dioxane / water / concentrated hydrochloric acid = 180 / 60 / 20 mL and reacted overnight at 110 °C. After the reaction was complete, the reaction solution was concentrated to dryness, diluted with water, extracted three times with ethyl acetate, dried over anhydrous sodium sulfate, and concentrated. The crude product was purified by column chromatography to give the title compound (12 g).

[0118]

[0119] MS(ESI)m / z(M+H) + =397.9.

[0120] Step 4: Preparation of 7-bromo-2,3-dihydroisoquinoline-4-one

[0121] 12 g of 3-(N-(3-bromophenyl)-4-methylphenylsulfonamide)propionic acid was dissolved in 100 mL of anhydrous dichloromethane. Thionyl chloride (10.8 mL) and 2-3 drops of N,N-dimethylformamide were added, and the mixture was refluxed at 40 °C for 2 h. After the starting material was consumed by LC-MS, the reaction solution was concentrated to dryness and diluted with 50 mL of anhydrous dichloromethane. 8 g of aluminum chloride was then dissolved in anhydrous dichloromethane, and the solution was added dropwise to the aluminum chloride solution in an ice bath. After the addition was complete, the reaction was allowed to proceed overnight at room temperature. After the starting material was consumed by TLC, the reaction was quenched with ice water, neutralized with sodium hydroxide solution, extracted three times with ethyl acetate, dried over anhydrous sodium sulfate, and concentrated. The crude product was purified by column chromatography to give the title compound (3.5 g).

[0122]

[0123] MS(ESI)m / z(M+H) + =225.9.

[0124] Step 5: Preparation of tert-butyl 7-bromo-4-oxo-3,4-dihydroquinoline-1(2H)-carboxylic acid

[0125] 7-Bromo-2,3-dihydroisoquinoline-4-one (762 mg) was dissolved in 20 mL of anhydrous dichloromethane. 4-Dimethylaminopyridine (830 mg) and di-tert-butyl dicarbonate (873 mg) were added under ice bath conditions, and the reaction was carried out at room temperature for 4 h. After the starting material was consumed by TLC, the reaction solution was concentrated to dryness. The crude product was purified by column chromatography to obtain the title compound (795 mg).

[0126]

[0127] MS(ESI)m / z(M+H) + =326.0.

[0128] Step 6: Preparation of tert-butyl 7-bromo-4-((tert-butylsulfinyl)imino)-3,4-dihydroquinoline-1(2H)-carboxylate

[0129] 7-Bromo-4-oxo-3,4-dihydroquinoline-1(2H)-carboxylic acid tert-butyl ester (795 mg) and tert-butylsulfinamide (443 mg) were dissolved in 2-methyltetrahydrofuran (10 mL), and 1 mL of tetraethyl titanate was added. The mixture was then transferred to 80 °C and reacted overnight. After the starting material was consumed by TLC, the mixture was diluted with ethyl acetate, 1 mL of water was added, filtered, concentrated, and the crude product was purified by column chromatography to give the title compound (907 mg).

[0130]

[0131] MS(ESI)m / z(M+H) + =429.1

[0132] Step 7: Preparation of tert-butyl-7-bromo-4-(1,1-dimethylethylsulfinamido)-3,4-dihydroquinoline-1(2H)-carboxylate

[0133] 907 mg of tert-butyl-7-bromo-4-((tert-butylsulfinyl)imino)-3,4-dihydroquinoline-1(2H)-carboxylate was dissolved in 10 mL of anhydrous tetrahydrofuran. 6.3 mL of tri-sec-butylborohydride was slowly added in an ice bath, and the reaction was carried out at room temperature for 2 h. After the starting material was consumed by TLC, the reaction solution was concentrated to dryness. The crude product was purified by column chromatography to give the title compound (380 mg).

[0134]

[0135] MS(ESI)m / z(M+Na) + =453.0.

[0136] Step 8: Preparation of tert-butyl 7-(diethoxyphosphoryl)-4-((R)-1,1-dimethylethylsulfinamido)-3,4-dihydroquinoline-1(2H)-carboxylate

[0137] 300 mg of tert-butyl-7-bromo-4-((R)-1,1-dimethylethylsulfinamido)-3,4-dihydroquinoline-1(2H)-carboxylate, 193 mg of diethyl phosphite, 64 mg of tris(dibenzylacetone)dipalladium, 77.6 mg of 1,1'-bis(diphenylphosphine)ferrocene (dppf, 77.6 mg), and 141 mg of triethylamine were dissolved in 10 mL of toluene. The mixture was reacted overnight at 125 °C under nitrogen protection. After the starting material was consumed by TLC, the mixture was diluted with ethyl acetate, filtered, concentrated, and the crude product was purified by pre-TLC to give the title compound (96 mg).

[0138]

[0139] MS(ESI)m / z(M+Na) + =511.0.

[0140] Step 9: Preparation of (4-amino-1,2,3,4-tetrahydroquinoline-7-yl)phosphonate

[0141] 96 mg of tert-butyl 7-(diethoxyphosphoryl)-4-(1,1-dimethylethylsulfinamido)-3,4-dihydroquinoline-1(2H)-carboxylate was added to concentrated hydrochloric acid (20 mL) and stirred overnight at 100 °C. After the starting material was consumed by LC-MS, the mixture was concentrated under reduced pressure and purified by pre-HPLC to obtain the title compound (22.6 mg).

[0142]

[0143] MS(ESI)m / z(M-16) + =211.9.

[0144] 1H NMR (400MHz, Deuterium Oxide)δ7.22(dd,J=7.7,3.8Hz,1H),7.05(td,J=15.3,13.0,9.6Hz,2H),4.45(d,J=4.3Hz,1H),3.24(dt,J=7.3,3.3Hz,2H),2.16-1.94(m,2H).

[0145] Example 4: Preparation of (5-amino-8-(2-hydroxyethyl)-5,6,7,8-tetrahydro-1,8-naphthid-2-yl)phosphonate

[0146] Step 1: Preparation of 3-((-tert-butylsulfinyl)amino)-3-(2,6-dichloropyridin-3-yl)propyl methanesulfonate

[0147] 200 mg of N-(1-(2,6-dichloropyridin-3-yl)-3-hydroxypropyl)-2-methylpropane-2-sulfinamide was dissolved in 4 mL of dichloromethane and placed at 0 °C. Triethylamine (0.17 mL) and methanesulfonyl chloride (48 μL) were added sequentially, and the reaction was carried out at 0 °C for 1.5 h. TLC showed that the starting material was completely consumed. The mixture was quenched with water, extracted twice with dichloromethane, dried over anhydrous sodium sulfate, and concentrated under vacuum to obtain the title compound (240 mg).

[0148]

[0149] MS(ESI)m / z(M+H) + =402.9

[0150] Step 2: Preparation of N-(7-chloro-1-(2-hydroxyethyl)-1,2,3,4-tetrahydro-1,8-naphthidin-4-yl)-2-methylpropane-2-sulfinamide

[0151] Weigh 240 mg of 3-((tert-butylsulfinyl)amino)-3-(2,6-dichloropyridin-3-yl)propyl methanesulfonate, dissolve it in 5 mL of N,N-dimethylformamide, add 109 mg of ethanolamine and 0.25 mL of triethylamine, and react at 70 °C for 1.5 h. TLC showed that the starting material was completely consumed. The mixture was quenched with water, extracted twice with ethyl acetate, washed twice with saturated brine, dried over anhydrous sodium sulfate, and purified by Prep-TLC to give the title compound (127 mg).

[0152]

[0153] MS(ESI)m / z(M+H) + =332.0.

[0154] Step 3: Preparation of N-(1-(2-((tert-butyldimethylsilyl)oxy)ethyl)-7-chloro-1,2,3,4-tetrahydro-1,8-naphthidin-4-yl)-2-methylpropane-2-sulfinamide

[0155] N-(7-chloro-1-(2-hydroxyethyl)-1,2,3,4-tetrahydro-1,8-naphthidin-4-yl)-2-methylpropane-2-sulfinamide (110 mg) was weighed and dissolved in dichloromethane (4 mL), and cooled to 0 °C. Imidazole (45 mg) and tert-butyldimethylchlorosilane (100 mg) were added, and the mixture was heated to room temperature for 2 h. TLC showed that the starting material was completely consumed. The mixture was quenched with water, extracted three times with ethyl acetate, dried over anhydrous sodium sulfate, and purified by Prep-TLC to give the title compound (139 mg).

[0156]

[0157] MS(ESI)m / z(M+H) + =446.2

[0158] Step 4: Preparation of diethyl (8-(2-((tert-butyldimethylsilyl)oxy)ethyl)-5-((tert-butylsulfinyl)amino)-5,6,7,8-tetrahydro-1,8-naphthidium-2-yl)phosphonate

[0159] N-(1-(2-((tert-butyldimethylsilyl)oxy)ethyl)-7-chloro-1,2,3,4-tetrahydro-1,8-naphthidin-4-yl)-2-methylpropane-2-sulfinamide (139 mg) was weighed and dissolved in toluene (4 mL). Tris(dibenzylacetone)dipalladium (29 mg), 1,1'-bis(diphenylphosphine)ferrocene (35 mg), triethylamine (95 mg), and diethyl phosphite (87 mg) were added sequentially. The mixture was purged with argon three times and reacted at 120 °C for 3 h. TLC showed that the starting material was completely consumed. The mixture was quenched with water, extracted twice with ethyl acetate, dried over anhydrous sodium sulfate, and purified by Prep-TLC to give the title compound (32 mg).

[0160]

[0161] MS(ESI)m / z(M+H) + =548.2

[0162] Step 5: Preparation of (5-amino-8-(2-hydroxyethyl)-5,6,7,8-tetrahydro-1,8-naphthid-2-yl)phosphonate

[0163] 32 mg of diethyl (8-(2-((tert-butyldimethylsilyl)oxy)ethyl)-5-((tert-butylsulfinyl)amino)-5,6,7,8-tetrahydro-1,8-naphthid-2-yl)phosphonate was dissolved in concentrated hydrochloric acid (4 mL) and reacted at 100 °C for 3 h. LCMS showed that the starting material was completely consumed. The mixture was concentrated under reduced pressure and purified by pre-HPLC to obtain the title compound (5.23 mg).

[0164]

[0165] MS(ESI)m / z(M+H) + =274.0

[0166] 1 H NMR (400MHz, Deuterium Oxide)δ7.87(dd,J=7.4,2.9Hz,1H),7.09(t,J=8.1Hz,1H),4.63(t,J=5.2Hz,1H),3.87(t,J=4 .4Hz,2H),3.75-3.57(m,4H),2.29(ddt,J=14.2,9.5,4.8Hz,1H),2.19(dq,J=15.0,5.4Hz,1H).

[0167] Example 5: Preparation of (5-amino-8-benzyl-5,6,7,8-tetrahydro-1,8-naphthid-2-yl) phosphonate

[0168]

[0169] Step 1: Preparation of 3-((tert-butylsulfinyl)amino)-3-(2,6-dichloropyridin-3-yl)propyl methanesulfonate

[0170] 250 mg of N-(1-(2,6-dichloropyridin-3-yl)-3-hydroxypropyl)-2-methylpropane-2-sulfinamide was dissolved in 4 mL of dichloromethane and placed at 0 °C. Triethylamine (0.21 mL) and methanesulfonyl chloride (60 μL) were added sequentially, and the reaction was carried out at 0 °C for 1.5 h. TLC showed that the starting material was completely consumed. The mixture was quenched with water, extracted twice with dichloromethane, dried over anhydrous sodium sulfate, and concentrated under vacuum to obtain the title compound (300 mg).

[0171]

[0172] MS(ESI)m / z(M+H) + =402.9

[0173] Step 2: Preparation of N-(1-benzyl-7-chloro-1,2,3,4-tetrahydro-1,8-naphthidin-4-yl)-2-methylpropane-2-sulfinamide

[0174] Weigh 300 mg of 3-((tert-butylsulfinyl)amino)-3-(2,6-dichloropyridin-3-yl)propyl methanesulfonate, dissolve it in N,N-dimethylformamide (6 mL), add benzylamine (160 mg) and triethylamine (0.30 mL), and react at 70 °C for 2 h. TLC showed that the starting material was completely consumed. Quench with water, extract twice with ethyl acetate, wash twice with saturated brine, dry with anhydrous sodium sulfate, and purify by column chromatography to obtain the title compound (356 mg).

[0175]

[0176] MS(ESI)m / z(M+H) + =378.0

[0177] Step 3: Preparation of diethyl (8-benzyl-5-((tert-butylsulfinyl)amino)-5,6,7,8-tetrahydro-1,8-naphthidium-2-yl)phosphonate

[0178] N-(1-benzyl-7-chloro-1,2,3,4-tetrahydro-1,8-naphthidin-4-yl)-2-methylpropane-2-sulfinamide (180 mg) was weighed and dissolved in toluene (2 mL). Tris(dibenzylacetone)dipalladium (44 mg), 1,1'-bis(diphenylphosphine)ferrocene (53 mg), triethylamine (0.2 mL), and diethyl phosphite (132 mg) were added sequentially. The mixture was purged with argon three times and reacted at 120 °C for 9 h. TLC showed that the starting material was completely consumed. The mixture was quenched with water, extracted twice with ethyl acetate, dried over anhydrous sodium sulfate, and purified by Prep-TLC to give the title compound (32 mg).

[0179]

[0180] MS(ESI)m / z(M+H) + =480.1

[0181] Step 4: Preparation of (5-amino-8-benzyl-5,6,7,8-tetrahydro-1,8-naphthid-2-yl)phosphonate

[0182] 27 mg of diethyl (8-benzyl-5-((tert-butylsulfinyl)amino)-5,6,7,8-tetrahydro-1,8-naphthid-2-yl)phosphonate was dissolved in concentrated hydrochloric acid (4 mL) and reacted at 100 °C for 3 h. LCMS showed that the starting material was completely consumed. The mixture was concentrated under reduced pressure and purified by pre-HPLC to obtain the title compound (3.44 mg).

[0183]

[0184] MS(ESI)m / z(M+H) + =320.2

[0185] 1 H NMR (400MHz, Deuterium Oxide)δ7.91(dd,J=6.9,2.7Hz,1H),7.33(q,J=7.5,7.0Hz,5H),7.09(t,J=8.2Hz,1H),4.95–4.80 (m,2H),4.67(s,1H),3.84–3.72(m,2H),2.35(tt,J=9.5,4.8Hz,1H),2.23(dq,J=15.1,5.4Hz,1H).

[0186] Example 6: Preparation of (5-amino-5,6,7,8-tetrahydro-1,8-naphthid-2-yl) phosphonate

[0187]

[0188] Step 1: Preparation of 3-((-tert-butylsulfinyl)amino)-3-(2,6-dichloropyridin-3-yl)propyl methanesulfonate

[0189] 270 mg of N-(1-(2,6-dichloropyridin-3-yl)-3-hydroxypropyl)-2-methylpropane-2-sulfinamide was dissolved in 5 mL of dichloromethane and placed at 0 °C. Triethylamine (0.21 mL) and methanesulfonyl chloride (64 μL) were added sequentially, and the reaction was carried out at 0 °C for 1.5 h. TLC showed that the starting material was completely consumed. The mixture was quenched with water, extracted twice with dichloromethane, dried over anhydrous sodium sulfate, and concentrated under vacuum to obtain the title compound (330 mg).

[0190]

[0191] MS(ESI)m / z(M+H) + =402.9

[0192] Step 2: Preparation of N-(7-chloro-1-(4-methoxybenzyl)-1,2,3,4-tetrahydro-1,8-naphthidin-4-yl)-2-methylpropane-2-sulfinamide

[0193] Weigh 330 mg of 3-((tert-butylsulfinyl)amino)-3-(2,6-dichloropyridin-3-yl)propyl methanesulfonate, dissolve it in N,N-dimethylformamide (6 mL), add p-methoxybenzylamine (338 mg) and triethylamine (0.34 mL), and react at 70 °C for 3 h. TLC showed that the starting material was completely consumed. Quench with water, extract twice with ethyl acetate, wash twice with saturated brine, dry with anhydrous sodium sulfate, and purify by column chromatography to obtain the title compound (336 mg).

[0194]

[0195] MS(ESI)m / z(M+H) + =408.1

[0196] Step 3: Preparation of diethyl (5-((tert-butylsulfinyl)amino)-8-(4-methoxybenzyl)-5,6,7,8-tetrahydro-1,8-naphthid-2-yl)phosphonate

[0197] 336 mg of N-(7-chloro-1-(4-methoxybenzyl)-1,2,3,4-tetrahydro-1,8-naphthidin-4-yl)-2-methylpropane-2-sulfinamide was weighed and dissolved in toluene (6 mL). Tris(dibenzylacetone)dipalladium (76 mg), 1,1'-bis(diphenylphosphine)ferrocene (92 mg), triethylamine (0.35 mL), and diethyl phosphite (0.21 mL) were added sequentially. The mixture was purged with argon three times and reacted overnight at 120 °C. TLC showed that the starting material was consumed. The mixture was quenched with water, extracted twice with ethyl acetate, dried over anhydrous sodium sulfate, and purified by Prep-TLC to give the title compound (110 mg).

[0198]

[0199] MS(ESI)m / z(M+H) + =510.2

[0200] Step 4: Preparation of (5-amino-5,6,7,8-tetrahydro-1,8-naphthid-2-yl)phosphonate

[0201] 65 mg of diethyl (5-((tert-butylsulfinyl)amino)-8-(4-methoxybenzyl)-5,6,7,8-tetrahydro-1,8-naphthid-2-yl)phosphonate was dissolved in concentrated hydrochloric acid (4 mL) and reacted overnight at 100 °C. LCMS showed that the starting material was completely consumed. The mixture was concentrated under reduced pressure and purified by pre-HPLC to obtain the title compound (17.5 mg).

[0202]

[0203] MS(ESI)m / z(M+H) + =230.1

[0204] 1 H NMR (400MHz, Deuterium Oxide)δ7.87(dd,J=7.4,2.8Hz,1H),7.02(dd,J=9.0,7.3Hz,1H),4.63(t,J=5.0Hz,1H),3.5 9(dt,J=13.9,5.2Hz,1H), 3.45(ddd,J=13.9,9.2,4.8Hz,1H), 2.15(dq,J=10.3,5.0Hz,2H).

[0205] Example 7: Preparation of (5-amino-5,6,7,8-tetrahydronaphth-2-yl)phosphonate

[0206]

[0207] Step 1: Preparation of N-(6-bromo-3,4-dihydronaphthalene-1(2H)-alkylene)-2-methylpropane-2-sulfinamide

[0208] 6-Bromo-3,4-dihydronaphthyl-1(2H)-one (300 mg, 1.33 mmol) and 2-methylpropane-2-sulfinamide (242 mg) were dissolved in 2-methylfuran and added to tetraethyl titanate (1 mL). The reaction was carried out at 85 °C for 4 h under nitrogen protection, and the reaction was monitored by TLC until complete. After cooling to room temperature, the reaction solution was added dropwise to a mixture of ethyl acetate and saturated brine. The mixture was filtered through a diatomaceous earth filter, and the filtrate was concentrated and purified by column chromatography to obtain the title compound (399 mg).

[0209]

[0210] MS(ESI)m / z(M+H) + =328.0 / 330.0.

[0211] Step 2: Preparation of N-(6-bromo-1,2,3,4-tetrahydronaphth-1-yl)-2-methylpropane-2-sulfinamide

[0212] N-(6-bromo-3,4-dihydronaphthyl-1(2H)-alkylene)-2-methylpropane-2-sulfinamide (399 mg) was added dropwise to a solution of tri-sec-butylborohydride (3.7 mL) under nitrogen protection at -15 °C. The reaction was carried out at this temperature for 1 h, and the reaction was monitored by TLC until complete. The reaction was quenched with saturated ammonium chloride solution, extracted with ethyl acetate, dried over anhydrous sodium sulfate, concentrated, and purified by column chromatography to give the title compound (316 mg).

[0213]

[0214] MS(ESI)m / z(M+H) + =330.0 / 332.0.

[0215] Step 3: Preparation of (5-((tert-butylsulfinyl)amino)diethyl)-5,6,7,8-tetrahydronaphth-2-yl)phosphonate

[0216] N-(6-bromo-1,2,3,4-tetrahydronaphth-1-yl)-2-methylpropane-2-sulfinamide (158 mg) was weighed and dissolved in toluene (5 mL). Diethyl phosphite (133 mg), triethylamine (200 μL), tris(dibenzylacetone)dipalladium (44 mg), and 1,1'-bis(diphenylphosphine)ferrocene (54 mg) were added sequentially. The mixture was purged with argon three times and reacted at 120 °C for 2 h. TLC showed that the starting material was completely consumed. The mixture was quenched with water, extracted twice with ethyl acetate, dried over anhydrous sodium sulfate, and purified by Prep-TLC to give the title compound (112 mg).

[0217]

[0218] MS(ESI)m / z(M+H) + =388.1.

[0219] Step 4: Preparation of (5-amino-5,6,7,8-tetrahydronaphth-2-yl)phosphonate

[0220] (5-((tert-butylsulfinyl)amino)diethyl)-5,6,7,8-tetrahydronaphth-2-yl)phosphonate (112 mg) was weighed and dissolved in concentrated hydrochloric acid (10 mL), and reacted overnight at 100 °C. LCMS showed that the starting material was completely consumed. The mixture was concentrated under reduced pressure and separated by pre-HPLC to obtain the title compound (30 mg).

[0221]

[0222] MS(ESI)m / z(MH) - =226.0.

[0223] 1 H NMR (400MHz, Deuterium Oxide) δ7.47(t,J=11.2Hz,2H),7.32(dd,J=7.8,3.4Hz,1H),4.50(t,J=5.4Hz,1H),2.77(dtd,J=24.6 ,17.2,6.5Hz,2H),2.08(dt,J=13.5,6.9Hz,1H),1.92(dq,J=14.2,5.2Hz,1H),1.79(p,J=6.7Hz,2H).

[0224] Example 8: Preparation of (5-amino-5,6,7,8-tetrahydroquinoline-2-yl)phosphonate

[0225]

[0226] Step 1: Preparation of N-(2-chloro-7,8-dihydroquinoline-5(6H)-alkylene)-2-methylpropane-2-sulfinamide

[0227] 300 mg of 2-chloro-7,8-dihydroquinoline-5(6H)-one and 300 mg of 2-methylpropane-2-sulfinamide were dissolved in 10 mL of 2-methylfuran and added to 1 mL of tetraethyl titanate. The reaction was carried out at 85 °C for 4 h under nitrogen protection, and the reaction was monitored by TLC until complete. After cooling to room temperature, the reaction solution was added dropwise to a mixture of ethyl acetate and saturated brine. The mixture was filtered through a diatomaceous earth filter, and the filtrate was concentrated and purified by column chromatography to obtain the title compound (433 mg).

[0228]

[0229] MS(ESI)m / z(M+H) + =285.9.

[0230] Step 2: Preparation of N-((S)-2-chloro-5,6,7,8-tetrahydroquinoline-5-yl)-2-methylpropane-2-sulfinamide

[0231] N-(2-chloro-7,8-dihydroquinoline-5(6H)-alkylene)-2-methylpropane-2-sulfinamide (433 mg) was added dropwise to a solution of tri-sec-butylborohydride (4.5 mL) under nitrogen protection at -15 °C. The reaction was carried out at this temperature for 1 h, and the reaction was monitored by TLC until complete. The reaction was quenched with saturated ammonium chloride solution, extracted with ethyl acetate, dried over anhydrous sodium sulfate, concentrated, and purified by column chromatography to give the title compound (276 mg).

[0232]

[0233] MS(ESI)m / z(M+H) + =287.0.

[0234] Step 3: Preparation of (5-((tert-butylsulfinyl)amino)diethyl)-5,6,7,8-tetrahydroquinoline-2-yl)phosphonate

[0235] 100 mg of N-(2-chloro-5,6,7,8-tetrahydroquinolin-5-yl)-2-methylpropane-2-sulfinamide was dissolved in 5 mL of toluene. Diethyl phosphite (97 mg), triethylamine (98 μL), tris(dibenzylacetone)dipalladium (32 mg), and 1,1'-bis(diphenylphosphine)ferrocene (39 mg) were added sequentially. The mixture was purged three times with argon gas and reacted at 120 °C for 2 h. TLC showed complete consumption of the starting material. The mixture was quenched with water, extracted twice with ethyl acetate, dried over anhydrous sodium sulfate, and purified by Prep-TLC to give the title compound (68 mg, 50% yield).

[0236]

[0237] MS(ESI)m / z(M+H) + =389.2.

[0238] Step 4: Preparation of (5-amino-5,6,7,8-tetrahydroquinoline-2-yl)phosphonate

[0239] (5-((tert-butylsulfinyl)amino)diethyl)-5,6,7,8-tetrahydroquinoline-2-yl)phosphonate (68 mg) was dissolved in concentrated hydrochloric acid (5 mL) and reacted overnight at 100 °C. LCMS showed that the starting material was completely consumed. The mixture was concentrated under reduced pressure and separated by pre-HPLC to obtain the title compound (25 mg).

[0240]

[0241] MS(ESI)m / z(M+H) + =229.0.

[0242] 1 H NMR (400MHz, Deuterium Oxide)δ8.49(dd,J=8.0,2.5Hz,1H),8.02(t,J=7.8Hz,1H),4.75(d,J=4.6Hz,1H),3.15(tdd,J =19.1,12.9,5.9Hz,2H),2.19(tt,J=12.2,6.2Hz,1H),2.00(ddq,J=11.3,7.9,4.9,4.1Hz,3H).

[0243] Example 9: Preparation of (4-aminothiopyran-7-yl)phosphonate

[0244]

[0245] Step 1: Preparation of 2-((3-bromophenyl)thio)propionic acid

[0246] 0.27 mL of m-bromothiophenol was dissolved in 5 mL of 2 M sodium hydroxide aqueous solution and reacted at 100 °C for 2 h. The reaction temperature was then lowered to 60 °C, and 3-chloropropionic acid (302 mg) was added. The reaction was continued overnight at 60 °C, and TLC was used to monitor the reaction until complete. After cooling to room temperature, the pH of the system was adjusted to 2 with 3 M hydrochloric acid aqueous solution in an ice bath. A large amount of solid precipitated. The solid was filtered, and the filter cake was washed three times with water. The filter cake was then removed and dried under reduced pressure to obtain the title compound (500 mg).

[0247]

[0248] MS(ESI)m / z(MH) - =258.8 / 260.8.

[0249] Step 2: Preparation of 7-bromothiobenzodihydropyran-4-one

[0250] 500 mg of 2-((3-bromophenyl)thio)propionic acid was dissolved in 10 mL of cold concentrated sulfuric acid. The mixture was stirred in an ice-water bath for half an hour, then moved to room temperature and stirred for 3 hours. The reaction was monitored by TLC until complete. The mixture was slowly poured into ice and extracted three times with ethyl acetate. The organic phases were combined and washed once with saturated sodium bicarbonate solution and once with saturated brine. The mixture was dried over anhydrous sodium sulfate, concentrated, and purified by column chromatography to give the title compound (320 mg).

[0251]

[0252] MS(ESI)m / z(M+H) + =242.9 / 244.8.

[0253] 1 H NMR(400MHz,Chloroform-d)δ7.98-7.96(d,J=8.5Hz,1H),7.48-7.47(d,J=1.9H z,1H),7.33-7.30(dd,J=8.5,1.9Hz,1H),3.28-3.25(m,2H),3.01-2.96(m,2H).

[0254] Step 3: Preparation of N-(7-bromothiopyran-4-alkylene)-2-methylpropane-2-sulfinamide

[0255] 320 mg of 7-bromothiobenzodihydropyran-4-one and 240 mg of 2-methylpropane-2-sulfinamide were dissolved in 3 mL of tetraethyl titanate. The reaction was carried out at 100 °C for 1 h under nitrogen protection, and the reaction was monitored by TLC until complete. After cooling to room temperature, the reaction solution was added dropwise to a mixture of ethyl acetate and saturated brine. The mixture was filtered through a diatomaceous earth filter, and the filtrate was concentrated and purified by column chromatography to obtain the title compound (320 mg).

[0256]

[0257] MS(ESI)m / z(M+H) + =345.9 / 348.0.

[0258] Step 4: Preparation of N-(7-bromothiobenzopyran-4-yl)-2-methylpropane-2-sulfinamide

[0259] N-(7-bromothiopyran-4-alkylene)-2-methylpropane-2-sulfinamide (320 mg) was added dropwise to a solution of tri-sec-butylborohydride (2.8 mL) under nitrogen protection at -15 °C. The reaction was carried out at this temperature for 1 h, and the reaction was monitored by TLC until complete. The reaction was quenched with saturated ammonium chloride solution, extracted with ethyl acetate, dried over anhydrous sodium sulfate, concentrated, and purified by column chromatography to give the title compound (260 mg).

[0260]

[0261] MS(ESI)m / z(M+H) + =369.8 / 371.9.

[0262] 1 H NMR (400MHz, Chloroform-d) δ7.29 (d, J=2.7Hz, 1H), 7.23-7.15 (m, 2H), 4.53-4.49 (ddd, J=9.1, 5.9, 3.6Hz, 1H), 3.49-3.47 (d, J= 8.7Hz,1H),3.22-3.15(ddd,J=12.9,10.5,3.6Hz,1H),3.06-3.00(ddd,J=12.8,6.4,3.9Hz,1H),2.48-2.32(m,2H),1.25(s,9H).

[0263] Step 5: Preparation of diethyl (4-((tert-butylsulfinyl)amino)thiopyran-7-yl)phosphonate

[0264] N-(7-bromothiobenzopyran-4-yl)-2-methylpropane-2-sulfinamide (50 mg) was dissolved in toluene (3 mL), followed by the addition of diethyl phosphite (39 mg), triethylamine (58 μL), tris(dibenzylacetone)dipalladium (13 mg), and 1,1'-bis(diphenylphosphine)ferrocene (16 mg). The mixture was purged with argon three times and reacted at 120 °C for 2 h. TLC showed that the starting material was completely consumed. The mixture was then quenched with water, extracted twice with ethyl acetate, dried over anhydrous sodium sulfate, and purified by Prep-TLC to obtain the title compound (50 mg).

[0265]

[0266] MS(ESI)m / z(M+H) + =406.1.

[0267] Step 6: Preparation of (4-aminothiopyran-7-yl)phosphonate

[0268] 50 mg of diethyl (4-((tert-butylsulfinyl)amino)thiopyran-7-yl)phosphonate was dissolved in 4 mL of concentrated hydrochloric acid and reacted overnight at 100 °C. LCMS showed that the starting material was completely consumed. The mixture was concentrated under reduced pressure and separated by pre-HPLC to obtain the title compound (4.1 mg).

[0269]

[0270] MS(ESI)m / z(MH) - =243.9.

[0271] 1 H NMR (400MHz, Deuterium Oxide)δ7.44-7.41(d,J=13.2Hz,1H),7.39-7.27(m,2H),4.60-4.58(t,J=4.3Hz,1H),3.12-2.97(m,2H),2.43-2.34(m,1H),2.28-2.20(m,1H).

[0272] Example 10: Preparation of (4-amino-3,4-dihydro-2H-thiopyrano[2,3-b]pyridin-7-yl)phosphonate hydrochloride

[0273] Step 1: Preparation of 3-((tert-butylsulfinyl)amino)-3-(2,6-dichloropyridin-3-yl)propyl methanesulfonate

[0274] 360 mg of N-(1-(2,6-dichloropyridin-3-yl)-3-hydroxypropyl)-2-methylpropane-2-sulfinamide was dissolved in 6 mL of dichloromethane and placed at 0 °C. Triethylamine (0.46 mL) and methanesulfonyl chloride (86 μL) were added sequentially, and the reaction was carried out at 0 °C for 1.5 h. TLC showed that the starting material was completely consumed. The mixture was quenched with water, extracted twice with dichloromethane, dried over anhydrous sodium sulfate, and concentrated under vacuum to obtain the title compound (440 mg).

[0275]

[0276] MS(ESI)m / z(M+H) + =402.9.

[0277] Step 2: Preparation of (-3-((tert-butylsulfinyl)amino)-3-(2,6-dichloropyridin-3-yl)propyl)ethanethiol ester

[0278] Weigh 200 mg of 3-((tert-butylsulfinyl)amino)-3-(2,6-dichloropyridin-3-yl)propyl methanesulfonate, dissolve it in 4 mL of N,N-dimethylformamide, and react with 57 mg of potassium thioacetate at 40 °C for 2 h. TLC showed that the starting material was completely consumed. The mixture was quenched with water, extracted twice with ethyl acetate, washed twice with saturated brine, dried over anhydrous sodium sulfate, and concentrated under vacuum to obtain the title compound (220 mg).

[0279]

[0280] MS(ESI)m / z 0(M+H) + =383..

[0281] Step 3: Preparation of N-(7-chloro-3,4-dihydro-2H-thiopyrano[2,3-b]pyridin-4-yl)-2-methylpropane-2-sulfinamide

[0282] (3-((tert-butylsulfinyl)amino)-3-(2,6-dichloropyridin-3-yl)propyl)ethanethiol ester (220 mg) was weighed and dissolved in tetrahydrofuran (4 mL). 4 mL of 2M sodium hydroxide aqueous solution was added, and the reaction was carried out at 60 °C for 2 h. TLC showed that the starting material was completely consumed. The mixture was quenched with water, extracted three times with ethyl acetate, dried over anhydrous sodium sulfate, and purified by column chromatography to obtain the title compound (108 mg).

[0283]

[0284] MS(ESI)m / z(M+H) + =305.0.

[0285] Step 4: Preparation of diethyl (4-((tert-butylsulfinyl)amino)-3,4-dihydro-2H-thiaro[2,3-b]pyridin-7-yl)phosphonate

[0286] 108 mg of N-(7-chloro-3,4-dihydro-2H-thiopyrano[2,3-b]pyridin-4-yl)-2-methylpropane-2-sulfinamide was dissolved in toluene (3 mL). Tris(dibenzylacetone)dipalladium (33 mg), 1,1'-bis(diphenylphosphine)ferrocene (40 mg), triethylamine (148 μL), and diethyl phosphite (92 μL) were added sequentially. The mixture was purged with argon three times and reacted at 120 °C for 6 h. TLC showed that the starting material was consumed. The mixture was quenched with water, extracted twice with ethyl acetate, dried over anhydrous sodium sulfate, and purified by Prep-TLC to give the title compound (45 mg).

[0287]

[0288] MS(ESI)m / z(M+H) + =407.1.

[0289] Step 5: Preparation of (4-amino-3,4-dihydro-2H-thiopyrano[2,3-b]pyridin-7-yl)phosphonate hydrochloride

[0290] 45 mg of diethyl (4-((tert-butylsulfinyl)amino)-3,4-dihydro-2H-thiaro[2,3-b]pyridin-7-yl)phosphonate was dissolved in concentrated hydrochloric acid (4 mL) and reacted overnight at 100 °C. LCMS showed that the starting material was completely consumed. The mixture was concentrated under reduced pressure and separated by pre-HPLC to obtain the title compound (5.49 mg).

[0291]

[0292] MS(ESI)m / z(M+H) + =247.0.

[0293] 1 H NMR (400MHz, Deuterium Oxide)δ7.87(dd,J=7.4,2.8Hz,1H),7.02(dd,J=9.0,7.3Hz,1H),4.63(t,J=5.0Hz,1H),3.5 9(dt,J=13.9,5.2Hz,1H), 3.45(ddd,J=13.9,9.2,4.8Hz,1H), 2.15(dq,J=10.3,5.0Hz,2H).

[0294] Example 11 Preparation of (5-amino-8-methyl-5,6,7,8-tetrahydro-1,8-naphthid-2-yl)phosphonate hydrochloride

[0295] Step 1: Preparation of 3-((tert-butylsulfinyl)amino)-3-(2,6-dichloropyridin-3-yl)propyl methanesulfonate

[0296] 360 mg of N-(1-(2,6-dichloropyridin-3-yl)-3-hydroxypropyl)-2-methylpropane-2-sulfinamide was dissolved in 6 mL of dichloromethane and placed at 0 °C. Triethylamine (0.46 mL) and methanesulfonyl chloride (86 μL) were added sequentially, and the reaction was carried out at 0 °C for 1.5 h. TLC showed that the starting material was completely consumed. The mixture was quenched with water, extracted twice with dichloromethane, dried over anhydrous sodium sulfate, and concentrated under vacuum to obtain the title compound (440 mg).

[0297]

[0298] MS(ESI)m / z(M+H) + =402.9.

[0299] Step 2: Preparation of N-(7-chloro-1-methyl-1,2,3,4-tetrahydro-1,8-naphthidin-4-yl)-2-methylpropane-2-sulfinamide

[0300] 220 mg of 3-((tert-butylsulfinyl)amino)-3-(2,6-dichloropyridin-3-yl)propyl methanesulfonate was weighed into a sealed tube, dissolved in 5 mL of tetrahydrofuran, followed by 1.64 mL of 2M tetrahydrofuran solution of methylamine and 0.23 mL of triethylamine. The reaction was carried out overnight at 100 °C. TLC showed that the starting material was completely consumed. The mixture was quenched with water, extracted twice with ethyl acetate, dried over anhydrous sodium sulfate, and purified by column chromatography to give the title compound (110 mg).

[0301]

[0302] MS(ESI)m / z(M+H) + =302.1.

[0303] Step 3: Preparation of diethyl (5-((tert-butylsulfinyl)amino)-8-methyl-5,6,7,8-tetrahydro-1,8-naphthidin-2-yl)phosphonate

[0304] N-(7-chloro-1-methyl-1,2,3,4-tetrahydro-1,8-naphthidin-4-yl)-2-methylpropane-2-sulfinamide (110 mg) was weighed and dissolved in toluene (3 mL). Tris(dibenzylacetone)dipalladium (34 mg, 0.037 mmol), 1,1'-bis(diphenylphosphine)ferrocene (41 mg), triethylamine (0.15 mL), and diethyl phosphite (94 μL) were added sequentially. The mixture was purged with argon three times and reacted at 120 °C for 6 h. TLC showed that the starting material was completely consumed. The mixture was quenched with water, extracted twice with ethyl acetate, dried over anhydrous sodium sulfate, and purified by Prep-TLC to give the title compound (73 mg).

[0305]

[0306] MS(ESI) m / z 404.1(M+H) + .

[0307] Step 4: Preparation of (5-amino-8-methyl-5,6,7,8-tetrahydro-1,8-naphthid-2-yl)phosphonate hydrochloride

[0308] (73 mg) of diethyl phosphonate (5-((tert-butylsulfinyl)amino)-8-methyl-5,6,7,8-tetrahydro-1,8-naphthid-2-yl)phosphonate was dissolved in concentrated hydrochloric acid (4 mL) and reacted at 100 °C for 8 h. LCMS showed that the starting material was completely consumed. The mixture was concentrated under reduced pressure and separated by pre-HPLC to obtain the title compound (26.7 mg).

[0309]

[0310] MS(ESI)m / z(M+H) + =244.0.

[0311] 1 H NMR (400MHz, Deuterium Oxide)δ7.82(dd,J=7.4,2.9Hz,1H),7.06(dd,J=9.3,7.2Hz,1H),4.62(t,J=5.1Hz,1H),3.66(dt,J=8.5,5.2Hz,2H),3.21(s,3H),2.31–2.11(m,2H).

[0312] Example 12 Preparation of (5-amino-8-methyl-5,6,7,8-tetrahydro-1,8-naphthid-2-yl)phosphonate hydrochloride

[0313]

[0314] Step 1: Preparation of 3-((tert-butylsulfinyl)amino)-3-(2,6-dichloropyridin-3-yl)propyl methanesulfonate

[0315] 500 mg of N-(1-(2,6-dichloropyridin-3-yl)-3-hydroxypropyl)-2-methylpropane-2-sulfinamide was dissolved in 8 mL of dichloromethane and placed at 0 °C. Triethylamine (0.64 mL) and methanesulfonyl chloride (119 μL) were added sequentially, and the reaction was carried out at 0 °C for 1.5 h. TLC showed that the starting material was completely consumed. The mixture was quenched with water, extracted twice with dichloromethane, dried over anhydrous sodium sulfate, and concentrated under vacuum to obtain the title compound (600 mg).

[0316]

[0317] MS(ESI)m / z(M+H) + =402.9

[0318] Step 2: Preparation of N-(7-chloro-1-isopropyl-1,2,3,4-tetrahydro-1,8-naphthidin-4-yl)-2-methylpropane-2-sulfinamide

[0319] 200 mg of 3-((tert-butylsulfinyl)amino)-3-(2,6-dichloropyridin-3-yl)propyl methanesulfonate was weighed into a sealed tube, dissolved in tetrahydrofuran (4 mL), followed by isopropylamine (127 μL) and triethylamine (0.20 mL). The reaction was carried out overnight at 120 °C. TLC showed that the starting material was completely consumed. The mixture was quenched with water, extracted twice with ethyl acetate, dried over anhydrous sodium sulfate, and purified by column chromatography to give the title compound (34 mg).

[0320]

[0321] MS(ESI)m / z(M+H) + =330.1.

[0322] Step 3: Preparation of diethyl (5-amino-8-isopropyl-5,6,7,8-tetrahydro-1,8-naphthid-2-yl)phosphonate

[0323] N-(7-chloro-1-isopropyl-1,2,3,4-tetrahydro-1,8-naphthidin-4-yl)-2-methylpropane-2-sulfinamide (34 mg) was weighed and dissolved in toluene (1.5 mL). Tris(dibenzylacetone)dipalladium (10 mg), 1,1'-bis(diphenylphosphine)ferrocene (12 mg), triethylamine (43 μL), and diethyl phosphite (40 μL) were added sequentially. The mixture was purged with argon three times and reacted overnight at 120 °C. TLC showed that the starting material was consumed. The mixture was concentrated under vacuum and purified by Prep-TLC to give the title compound (18 mg).

[0324]

[0325] MS(ESI)m / z(M+H) + =328.1.

[0326] Step 4: Preparation of (5-amino-8-methyl-5,6,7,8-tetrahydro-1,8-naphthid-2-yl)phosphonate hydrochloride

[0327] 18 mg of diethyl phosphonate (5-amino-8-isopropyl-5,6,7,8-tetrahydro-1,8-naphthid-2-yl)phosphonate was dissolved in concentrated hydrochloric acid (4 mL) and reacted overnight at 100 °C. LCMS showed that the starting material was completely consumed. The mixture was concentrated under reduced pressure and separated by pre-HPLC to obtain the title compound (5.2 mg).

[0328]

[0329] MS(ESI)m / z(M+H) + =272.1.

[0330] 1 H NMR (400MHz, Deuterium Oxide)δ7.77(dd,J=7.4,2.8Hz,1H),6.96(t,J=7.5Hz,1H),4.57(s,1H),4.43(p,J=6.6Hz,1H),3.63(dt,J=1 3.8, 5.1Hz, 1H), 3.45 (ddd, J=14.1, 8.4, 5.9Hz, 1H), 2.16 (q, J=5.2, 4.3Hz, 2H), 1.24 (dd, J=8.7, 6.6Hz, 6H).

[0331] Example 13

[0332]

[0333] Step 1: Preparation of 3-((tert-butylsulfinyl)amino)-3-(2,6-dichloropyridin-3-yl)propyl methanesulfonate

[0334] 300 mg of N-(1-(2,6-dichloropyridin-3-yl)-3-hydroxypropyl)-2-methylpropane-2-sulfinamide was dissolved in 5 mL of dichloromethane and placed at 0 °C. Triethylamine (0.38 mL) and methanesulfonyl chloride (71 μL) were added sequentially, and the reaction was carried out at 0 °C for 1.5 h. TLC showed that the starting material was completely consumed. The mixture was quenched with water, extracted twice with dichloromethane, dried over anhydrous sodium sulfate, and concentrated under vacuum to obtain the title compound (372 mg).

[0335]

[0336] MS(ESI) m / z 402.9(M+H) +

[0337] Step 2: Preparation of N-(7-chloro-1-isobutyl-1,2,3,4-tetrahydro-1,8-naphthidin-4-yl)-2-methylpropane-2-sulfinamide

[0338] 186 mg of 3-((tert-butylsulfinyl)amino)-3-(2,6-dichloropyridin-3-yl)propyl methanesulfonate was weighed into a sealed tube, dissolved in 4 mL of N,N-dimethylformamide, followed by 0.14 mL of isobutylamine and 0.19 mL of triethylamine. The reaction was carried out at 70 °C for 3 h. TLC showed that the starting material was completely consumed. The mixture was quenched with water, extracted twice with ethyl acetate, dried over anhydrous sodium sulfate, and purified by column chromatography to obtain the title compound (70 mg).

[0339]

[0340] MS(ESI)m / z(M+H) + =344.1.

[0341] Step 3: Preparation of diethyl (5-((tert-butylsulfinyl)amino)-8-isobutyl-5,6,7,8-tetrahydro-1,8-naphthidin-2-yl)phosphonate

[0342] 70 mg of N-(7-chloro-1-isobutyl-1,2,3,4-tetrahydro-1,8-naphthidin-4-yl)-2-methylpropane-2-sulfinamide was dissolved in toluene (2 mL). Tris(dibenzylacetone)dipalladium (19 mg), 1,1'-bis(diphenylphosphine)ferrocene (23 mg), triethylamine (85 μL), and diethyl phosphite (52 μL) were added sequentially. The mixture was purged three times with argon gas and reacted at 120 °C for 10 h. TLC showed that the starting material was completely consumed. The mixture was quenched with water, extracted twice with ethyl acetate, dried over anhydrous sodium sulfate, and purified by Prep-TLC to obtain the title compound (20 mg).

[0343]

[0344] MS(ESI)m / z(M+H) + =446.1.

[0345] Step 4: Preparation of (5-amino-8-isobutyl-5,6,7,8-tetrahydro-1,8-naphthid-2-yl)phosphonate hydrochloride

[0346] 20 mg of diethyl (5-((tert-butylsulfinyl)amino)-8-isobutyl-5,6,7,8-tetrahydro-1,8-naphthid-2-yl)phosphonate was dissolved in 4 mL of concentrated hydrochloric acid and reacted overnight at 100 °C. LCMS showed that the starting material was completely consumed. The mixture was concentrated under reduced pressure, and the title compound (12 mg) was separated by pre-HPLC.

[0347]

[0348] MS(ESI)m / z(M+H) + =286.1

[0349] 1 H NMR (400MHz, Deuterium Oxide)δ7.82(dd,J=7.5,2.7Hz,1H),6.99(t,J=7.6Hz,1H),4.62(t,J=5.1Hz,1H),3.65(tdd,J=17.9,10.1,4.6Hz ,2H),3.49(dd,J=15.2,7.5Hz,1H),3.34(dd,J=15.3,7.9Hz,1H),2.32–2.04(m,3H),0.91(dd,J=6.7,1.6Hz,6H).

[0350] Using raw materials and reagents purchased through conventional commercial channels, and referring to the preparation method of the foregoing embodiments combined with conventional separation and purification methods in the art, the compounds of the following embodiments were prepared:

[0351]

[0352]

[0353]

[0354]

[0355]

[0356] Example 33

[0357]

[0358] Step 1: Preparation of tert-butyl 7-bromo-4-oxo-2,3-dihydroisoquinoline-1(2H)-carboxylic acid

[0359] 7-Bromo-2,3-dihydroisoquinoline-4-one (762 mg) was dissolved in 20 mL of anhydrous dichloromethane. 4-Dimethylaminopyridine (830 mg) and di-tert-butyl dicarbonate (873 mg) were added under ice bath conditions, and the reaction was carried out at room temperature for 4 h. After the starting material was consumed by TLC, the reaction solution was concentrated to dryness. The crude product was purified by column chromatography to obtain the title compound (795 mg).

[0360]

[0361] MS(ESI)m / z(M+H) + =326.0.

[0362] Step 2: Preparation of tert-butyl7-bromo-4-((tert-butylsulfinyl)imino)-3,4-dihydroisoquinoline-1(2H)-carboxylate

[0363] 7-Bromo-4-oxo-3,4-dihydroisoquinoline-1(2H)-carboxylic acid tert-butyl ester (795 mg) and tert-butylsulfinamide (443 mg) were dissolved in 2-methyltetrahydrofuran (10 mL), and 1 mL of tetraethyl titanate was added. The mixture was then transferred to 80 °C and reacted overnight. After the starting material was consumed by TLC, the mixture was diluted with ethyl acetate, 1 mL of water was added, filtered, concentrated, and the crude product was purified by column chromatography to give the title compound (907 mg).

[0364]

[0365] MS(ESI)m / z(M+H) + =429.1

[0366] Step 3: Preparation of tert-butyl-7-bromo-4-(1,1-dimethylethylsulfinamido)-3,4-dihydroisoquinoline-1(2H)-carboxylate

[0367] 907 mg of tert-butyl-7-bromo-4-((tert-butylsulfinyl)imino)-3,4-dihydroisoquinoline-1(2H)-carboxylate was dissolved in 10 mL of anhydrous tetrahydrofuran. 6.3 mL of tri-sec-butylborohydride was slowly added in an ice bath, and the reaction was carried out at room temperature for 2 h. After the starting material was consumed by TLC, the reaction solution was concentrated to dryness. The crude product was purified by column chromatography to give the title compound (380 mg).

[0368]

[0369] MS(ESI)m / z(M+Na) + =453.0.

[0370] Step 4: Preparation of tert-butyl 7-(diethoxyphosphoryl)-4-((R)-1,1-dimethylethylsulfinamido)-3,4-dihydroisoquinoline-1(2H)-carboxylate

[0371] 300 mg of tert-butyl-7-bromo-4-((R)-1,1-dimethylethylsulfinamido)-3,4-dihydroisoquinoline-1(2H)-carboxylate, 193 mg of diethyl phosphite, 64 mg of tris(dibenzylacetone)dipalladium, 77.6 mg of 1,1'-bis(diphenylphosphine)ferrocene (dppf, 77.6 mg), and 141 mg of triethylamine were dissolved in 10 mL of toluene. The mixture was reacted overnight at 125 °C under nitrogen protection. After the starting material was consumed by TLC, the mixture was diluted with ethyl acetate, filtered, concentrated, and the crude product was purified by pre-TLC to give the title compound (96 mg).

[0372]

[0373] MS(ESI)m / z(M+Na) + =511.0.

[0374] Step 5: Preparation of (4-amino-1,2,3,4-tetrahydroquinoline-7-yl)phosphonate

[0375] 96 mg of tert-butyl 7-(diethoxyphosphoryl)-4-(1,1-dimethylethylsulfinamido)-3,4-dihydroisoquinoline-1(2H)-carboxylate was added to concentrated hydrochloric acid (20 mL) and stirred overnight at 100 °C. After the starting material was consumed by LC-MS, the mixture was concentrated under reduced pressure and purified by pre-HPLC to obtain the title compound (22.6 mg).

[0376]

[0377] MS(ESI)m / z(M-16) + =211.9.

[0378] 1 H NMR (400MHz, Deuterium Oxide): δ7.74–7.63(m,1H),7.58–7.45(m,2H),4.96(t,J=5.8Hz,1H),4.58–4.33(m,2H),3.88(dd,J=13.8,5.8Hz,1H),3.66(dd,J=13.8,5.9Hz,1H).

[0379] Biological experiments

[0380] Test Example 1: Plasma Clot Degradation Experiment

[0381] 1. Experimental Objective

[0382] The inhibitory effect of the compounds of this invention on the degradation of human plasma clots was determined.

[0383] 2. Experimental Materials and Instruments

[0384]

[0385] 3. Experimental Procedure

[0386] 3.1 Collect fresh blood from healthy individuals, use 0.109M trisodium citrate as an anticoagulant, mix 1 part anticoagulant with 9 parts blood, centrifuge at 2000xg for 20 minutes at room temperature, collect the supernatant (i.e., plasma), aliquot and store at -80℃ for later use.

[0387] 3.2 On the day of the experiment, the plasma was thawed in a water bath at 37°C, and all reagents except tPA were preheated at 37°C.

[0388] 3.3 Add 12.5 μL of 80 mM CaCl2 (HEPES buffer, pH 7.4) to a 96-well plate, then add 25 μL of the test compound diluted with physiological saline at different concentrations. Add an equal volume of physiological saline to the negative control wells.

[0389] 3.4 Mix 50 μL of preheated plasma with 12.5 μL of 4 nM tPA (HEPES buffer, pH 7.4) and immediately add to a 96-well plate. Detect the absorbance at 405 nm, reading every 2 minutes for 15 hours.

[0390] 3.5 The absorbance value changes over time, first increasing and then decreasing. The time corresponding to the median absorbance value during the decreasing phase minus the time corresponding to the median absorbance value during the increasing phase is the plasma clot lysis time. Using the plasma clot lysis time of the negative control well as a reference, the inhibition rate was calculated relative to the plasma clot lysis time in wells with different compound concentrations.

[0391] Inhibition rate % = (1 - negative control well) Clot lysis time / compound pores Clot lysis time 100%

[0392] 3.6 Fitting dose-response curve

[0393] Plotting the logarithmic value of compound concentration on the X-axis and the percentage inhibition rate on the Y-axis, dose-response curves were fitted using the log(inhibitor) vs. response-variable slope method in GraphPadPrism 5 to derive the IC50 of each compound on cell activity. 50 value.

[0394] Calculation formula: Y = min + (max - min) / (1 + 10^(LogIC)) 50 -X)×Hillslope).

[0395] The inhibitory effect of the compounds of this invention on plasma clot degradation was determined through the above experiments, and the IC50 of the compounds in the embodiments of this invention was calculated. 50 The values ​​were all significantly lower than those of tranexamic acid, the most widely used hemostatic drug in clinical practice (IC50 under the same experimental conditions). 50 (4.75 μM).

[0396] IC of the compound in the embodiments of the present invention 50 ratio value (IC) 50 ratio = IC of the compound of this invention 50 Value / Chanamidocyanine IC 50 The values ​​are as follows:

[0397] 1 0.06 18 / 2 0.11 19 0.11 3 0.64 20 0.49 4 0.51 21 / 5 0.39 22 0.95 6 0.22 23 0.15 7 0.54 24 0.73 8 0.48 25 0.73 9 0.62 26 / 10 0.37 27 0.33 11 0.53 28 / 12 0.82 29 / 13 0.28 30 0.97 14 0.67 31 / 15 0.24 32 0.3 16 0.42 33 2.19 17 /

[0398] Note: " / " indicates that it was not detected.

[0399] Experimental data show that the compound of the present invention can effectively inhibit the degradation of plasma clots, has excellent coagulation and hemostatic activity, and its effective dose is far lower than that of the most frequently used hemostatic drugs in clinical practice. It can effectively avoid adverse reactions and complications caused by high-dose medication and has excellent prospects for drug development.

Claims

1. The pharmaceutically acceptable salt of the compound represented by Formula I: Formula I in, X is selected from CH2, NR3, O, and S; Y is selected from CH and N. R1 is selected from hydroxyl groups; R2 is selected from hydrogen; R3 is selected from hydrogen, C1-C4 alkyl, C1-C4 haloalkyl, C3-C6 cycloalkyl, 4-8 membered alicyclic group, -(CH2)mNRcRd, -Q-(R Q ) n ; Where m = 1, 2, 3, 4, and Rc and Rd are each independently selected from hydrogen, methyl, ethyl, propyl, isopropyl, n-butyl, and tert-butyl. Q is selected from C1-C4 alkylene groups, n is 1, and R... Q Selected from phenyl, tetrahydropyranyl, morpholinyl, piperidinyl, methylpiperidinyl, o-chlorophenyl, carboxyl, hydroxyl, dimethylamino, and halogen.

2. The compound according to claim 1, wherein its pharmaceutically acceptable salt is characterized in that, The alkyl group is selected from methyl, ethyl, propyl, isopropyl, n-butyl, and tert-butyl. The cycloalkyl group is selected from cyclopropyl, cyclobutyl, cyclopentyl, and cyclohexyl; The alicyclic group is selected from oxabutanyl, pyrrolidinyl, tetrahydrofuranyl, tetrahydropyranyl, tetrahydrothiophenyl, piperidinyl, and morpholinyl.

3. A compound having the following structure: 、 。 4. A pharmaceutical composition comprising at least one compound according to any one of claims 1-3, or a pharmaceutically acceptable salt thereof, and at least one pharmaceutically acceptable excipient.

5. Use of the compound of any one of claims 1-3 or a pharmaceutically acceptable salt thereof or the pharmaceutical composition of claim 4 for the preparation of a medicament having therapeutic activities of coagulation and hemostasis.

6. The use according to claim 5, characterized in that, The drug is used for abnormal bleeding caused by hyperfibrinolysis, as well as bleeding during and after surgery.

Images