CN105884755A - Deuterium-modified carbon glycoside derivate - Google Patents

Abstract

The invention belongs to the field of medinal chemistry and relates to a deuterium-modified carbon glycoside derivate, in particular to a compound shown in the formula I. The invention further relates to a preparation method of the deuterium-modified carbon glycoside derivate, a medicine composition comprising the deuterium-modified carbon glycoside derivate and application of the deuterium-modified carbon glycoside derivate or the medicine composition thereof for preparing medicine for treating diabetes.

Description

The carbon glycoside derivative that deuterium is modified

Technical field

The invention belongs to medicinal chemistry art, relate to the carbon glycoside derivative that deuterium is modified.The invention still further relates to its preparation method, The pharmaceutical composition of the carbon glycoside derivative containing deuterated modification, the carbon glycoside derivative of deuterated modification and pharmaceutical composition thereof are controlled in preparation Treat the purposes in the medicine of diabetes.

Background technology

SGLT2 is the novel targets of the treatment diabetes of discovered in recent years.Glucose in blood plasma is filtered in glomerulus, And be actively absorbed from the gastrointestinal tract again at Renal proximal tubular and to return in blood, research shows, SGLT2 is control this heavy absorption process main Albumen.Therefore, the activity of suppression SGLT2 just can increase the excretion of glucose in urine thus reduce blood sugar concentration. Canagliflozin (canagliflozin, (1S)-1,5-dehydrogenation-1-C-[3-[[5-(4-fluorophenyl)-2-thienyl] methyl]-4- Aminomethyl phenyl]-D-Glucose alcohol) be current first acquisition FDA approval SGLT2 inhibitor class medicine.Canagliflozin leads to Cross the suppression kidney heavily absorption to glucose, increase glucose excretion, and then reduce the blood sugar level that diabetic has raised. At present, canagliflozin single therapy and (include melbine, sulfonylurea, Pioglitazone and pancreas with other antidiabetic drug Island element) correlative study of drug combination is complete.Canagliflozin should not be used in type 1 diabetes patient, Diabetic ketosis Acid poisoning patient, severe renal impairment, End-stage renal disease patient or dialysis patient.

FDA requires that canagliflozin studies after improving five listings: cardiovascular lapse to test, in terms of pharmacovigilance Research (is strengthened malignant tumour, serious pancreatitis, severe allergic reaction, light sensitivity reaction, liver exception and Disadvantage pregnancy result Monitoring), bone security study and two researchs for child patient (include pharmacokinetics, pharmacodynamic study and safety Study on the efficiency).Side effect common for canagliflozin has Vaginal Fungi to infect (vulvovaginal candidiasis) and urinary tract sense Dye.Owing to canagliflozin is relevant to diuresis, so this medicine can cause blood volume to reduce, so cause orthostatic or Postural hypotension (when standing, blood pressure reduces suddenly), or therefore cause dizziness or faintness symptom.These symptoms are at treatment head Three months more typically.Although canagliflozin clinical efficacy is notable, but when continuing to need few side effects and effect clinically Between long highly effective and safe antidiabetic drug.

Summary of the invention

One aspect of the present invention offer compound shown in a kind of Formulas I or its pharmaceutically acceptable salt:

Wherein R₁、R₂Independently selected from H (hydrogen), D (deuterium) or OH；R₄、R₅、R₆、R₇、R₈、R₉、R₁₀、R₁₁、R₁₂、R₁₃、R₁₄、R₁₅ Separately selected from H (hydrogen) or D (deuterium), R₃Independently selected from C₁-C₄Alkyl or deuterated C₁-C₄Alkyl, condition is R₁～R₁₅In Including at least a D-atom.

In the certain preferred embodiments of the present invention, R₃Independently selected from CH₃、CH₂D、CHD₂Or CD₃。

In the certain preferred embodiments of the present invention, the example of compound shown in Formulas I is as follows:

In the embodiment of present invention further optimization, the example of the compound shown in Formulas I is as follows:

Term " pharmaceutically acceptable salt " refers to remain the biological efficacy of the free bronsted lowry acids and bases bronsted lowry of specific compound and do not have There is the salt of biology ill-effect.The example of pharmaceutically acceptable salt includes but not limited to: (1) acid-addition salts, and inorganic acid The salt that such as hydrochloric acid, sulfuric acid, hydrobromic acid, nitric acid, phosphoric acid etc. are formed；Or and organic acids such as malic acid, fumaric acid, maleic acid, Benzoic acid, phenylacetic acid, butanedioic acid, tartaric acid, citric acid, methanesulfonic acid, ethyl sulfonic acid, hydroxyacetic acid, cinnamic acid, pyruvic acid, formic acid, The salt that acetic acid, propionic acid, oxalic acid, malonic acid, acrylic acid, mandelic acid etc. are formed；Or (2) base addition salts, and alkali metal such as lithium, The salt that sodium, potassium etc. are formed；The salt formed with alkaline-earth metal such as calcium, magnesium etc.；With organic bases such as ammonium, choline, diethanol amine, rely Propylhomoserin, ethylenediamine, tert-butylamine, t-octanylamine, three (methylol) aminomethane, N-methyl glucose osamine, triethanolamine, dehydrogenated rosin The salt that amine etc. are formed.

If any atom of the compound of institute's labelled synthesis does not specify in the present invention, any of this atom can be represented A kind of stable isotope.Unless stated otherwise, when position a certain in structure is defined as H i.e. hydrogen (H-1), this position only contains Naturally occurring isotopic mass.Equally, unless stated otherwise, when position a certain in structure is defined as D i.e. deuterium (H-2), should Position containing at least big than naturally occurring isotopic mass (0.015%) 3340 times of isotopic mass (i.e. to I haven't seen you for ages 50.1% deuterium coordination Element).

In the present invention, the deuterated rate of the compound of institute's labelled synthesis refers to the isotopic content of labelled synthesis and naturally occurs The ratio of isotopic mass.In the present invention, the deuterated rate of each appointment D-atom of the compound of institute's labelled synthesis can be at least 3500 times (52.5%), at least 4000 times (60%), at least 4500 times (67.5%), at least 5000 times (75%), extremely It is 5500 times (82.5%), at least 6000 times (90%), at least 6333.3 times (95%) less, is at least 6466.7 times (97%), it is at least 6566.7 times (98.5%), at least 6600 times (99%), is at least 6633.3 times (99.5%).

Coordination ferritic (isotopologues) in the present invention refers to only have in terms of chemical constitution in isotopics not Same compound.In the present invention, the compound of institute's labelled synthesis has identical chemical constitution, only atom at its molecule and forms In isotopic change.Therefore, in the present invention institute's labelled synthesis can contain considerably less at ad-hoc location too containing deuteride The hydrogen isotope body of this position, in the present invention, the hydrogen isotope body of certain position in the compound of institute's labelled synthesis measures certainly Many factors, including deuterated reagent (D₂O、D₂、NaBD₄、LiAlD₄Deng) deuterium isotopic purity and introduce deuterium isotope The validity of synthetic method.But, the amount sum of the hydrogen isotope body of certain position the most this will be less than 49.9%.This The amount sum of the hydrogen isotope body of certain position in the compound of bright middle institute labelled synthesis will less than 47.5%, 40%, 32.5%, 25%, 17.5%, 10%, 5%, 3%, 1% or 0.5%.

In the present invention, the compound of institute's labelled synthesis has dissimilar chiral centres, asymmetric including caused by deuterium replacement Chiral centre.In the present invention, the compound of institute's labelled synthesis can be single enantiomer, it is also possible to be mixture of enantiomers.Cause This, in the present invention, the compound of institute's labelled synthesis may be divided with the form of the form of raceme or single corresponding isomers Separate out.Single corresponding isomers contains other isomers very small amount of, e.g., less than 25%, 10%, 2% or 1%.

Herein, any each atom being not designated as deuterium exists with its natural isotopic abundance.

Further aspect of the present invention provides a kind of method of compound prepared as shown in I, including:

A) at reducing agent (such as LiAlH₄、Et₃SiH、NaBH₄/AlCl₃、NaBD₄/AlCl₃In the presence of), Formula II compound It is reduced compound of Formula III；

B) in the presence of alkali (such as n-BuLi), formula III compound reacts with formula IV compound, obtains Formula V compound；

C) at reducing agent (such as Et₃SiH/BF₃-Et₂O、Et₃SiD/BF₃-Et₂O), in the presence of, Formula V compound is reduced into Compound of formula I.

Formula II compound can be prepared by following method:

Wherein, deuterated 2-bromothiophene can be bought from companies such as Aldrich；Also Pd-Pt-NaBH can be passed through₄Combination catalysis Prepared by the H-D exchange reaction of agent or phosphoric acid catalyzed；Also in the presence of bromide reagent NBS, deuterated thiophene can be changed into 2-bromine deuterium For thiophene.In the presence of palladium chtalyst reagent, Formula IV compound and Formula VII compound occur C-C coupling reaction to prepare Formula VIII Compound, the most deuterated or non-deuterated 4-fluorobenzoic boric acid VII can buy from companies such as Combiphos；Deuterated 4-fluorobenzoic boric acid VII Also (Butters, Mike et al, Angewandte Chemie, International can be prepared by literature method Edition,49(30),5156-5160,S5156/1-S5156/68；2010).In Lews acid such as AlCl₃、ZnO、ZnCl₂Deng examination In the presence of agent, Formula VIII compound and Formula IX compound generation Hooke acylation reaction prepare Formula II compound.

Present invention compound shown in formula I also can be prepared as follows,

Formula X compound can react generation grignard reagent in the presence of magnesium metal, then presses document report with Formula X I Method (such as Good, James A.D.et al, Journal of Medicinal Chemistry, 56 (5), 1878-1893； 2013) reaction obtains Formula II compound, and it can be further at reducing agent (such as NaBH₄Or NaBD₄In the presence of), reaction obtains formula XII compound, then replaces with trim,ethylchlorosilane and obtains Formula X III compound.In the presence of n-BuLi, by Formula X III chemical combination After thing is transformed into lithium reagent, forms Formula X IV compound with formula IV compound condensation, finally going back original reagent (such as Et₃SiH (D)/BF₃-Et₂O), in the presence of, reaction obtains compound of formula I.

Further aspect of the present invention provides a kind of pharmaceutical composition, and it contains the compound shown in Formulas I or it pharmaceutically can connect The salt being subject to, and one or more pharmaceutically acceptable carriers, excipient and/or medium.

" pharmaceutical composition " refers to compound or its salt and the usual acceptance in the art of one or more present invention For bioactive compound is delivered to organism, such as people, the preparation of carrier, excipient and/or medium.Drug regimen The purpose of thing is advantageous for giving organism the compound of the present invention.

Term " pharmaceutically acceptable carrier " refers to organism without obvious stimulation effect, and without compromising on this activity The biologically active of compound and those carriers of performance and diluent." pharmaceutically acceptable excipient and/or medium " refers to The inert substance of administration that be together administered with active component, beneficially active component." pharmaceutically acceptable carrier, figuration Agent and/or medium " include but not limited to can be used for people or any carrier of livestock animals, excipient, medium, glidant, sweetened Agent, diluent, preservative, dyestuff/colouring agent, flavoring reinforcing agent, surfactant, wetting agent, dispersant, disintegrant, suspending Agent, stabilizer, isotonic agent, solvent or emulsifying agent.The limiting examples of described excipient includes calcium carbonate, calcium phosphate, various Sugared and each kind of starch, cellulose derivative, gelatin, vegetable oil and polyethylene glycol etc..

In a pure form or with the compounds of this invention of suitable pharmaceutical compositions or its pharmaceutically acceptable salt Administration can carry out by providing any acceptable mode of administration of medicament of similar applications.The pharmaceutical composition of the present invention can By the compound of the present invention is prepared with suitable pharmaceutically acceptable supporting agent, diluent or excipient composition, and can It is configured to solid-state, semisolid, liquid or gaseous state preparation, such as tablet, pill, capsule, pulvis, granule, paste, emulsion, hangs Floating agent, solution, suppository, injection, inhalant, gel, microballoon and aerosol etc..

The classical pathway giving the compounds of this invention or its pharmaceutically acceptable salt or its pharmaceutical composition include but not Be limited to be administered orally, rectum, saturating mucous membrane, through enteral administration, or local, percutaneous, suction, parenteral, sublingual, vagina, intranasal, eye In, intraperitoneal, intramuscular, subcutaneous, intravenous administration.Preferably method of administration is oral administration.

The pharmaceutical composition of the present invention can use method manufacture well-known in the art, such as conventional mixing method, molten Solution, granulation, dragee method processed, levigate method, emulsion process, freeze-drying etc..

In preferred embodiments, pharmaceutical composition is oral form.For oral administration, can be by activating Compound mixes with pharmaceutically acceptable carrier well known in the art, excipient and/or medium, prepares this pharmaceutical composition.This A little carriers, excipient and medium can make the compound of the present invention be formulated into tablet, pill, lozenge, sugar-coat agent, capsule, liquid Body, gel, slurry agent, suspending agent etc., for the oral administration to patient.

Solid oral composition can be prepared by conventional mixing, filling or tabletting method.Such as, can be by following Method obtains: mixed with solid excipient by described reactive compound, the mixture of gained of optionally milling, if necessary Add other suitable assistant agent, then this mixture is processed into particle, obtain tablet or the core of sugar-coat agent.The tax being suitable for Shape agent includes but not limited to: filler, such as sugar, including lactose, sucrose, mannitol or D-sorbite；Cellulose preparation is the beautifulst Rice starch, wheaten starch, rice starch and farina；And other material, such as gelatin, tragacanth, methylcellulose, hydroxyl Propyl methocel, sodium cellulose glycolate and/or polyvinylpyrrolidone；Disintegrant, as PVPP, Agar or alginic acid, it is possible to use salt, such as mosanom.Can be according to method known in usual medicinal practice optionally to sugar-coat The core of agent is coated, and especially uses enteric coating.

Pharmaceutical composition could be applicable to parenteral, as the sterile solution agent of suitable unit dosage forms, supensoid agent or Freeze-drying prods.Suitable excipient, such as filler, buffer or surfactant can be used.

The dosage of the compound shown in formula I can be (such as age, body weight, to be treated according to the health of patient The type of disease and seriousness) and method of administration and change, and generally at about 0.01-300mg/kg/ days, clinician can basis Actual conditions eligibly select.

Further aspect of the present invention provides the compound shown in Formulas I or its salt pharmaceutically accepted to benefit from preparation treatment Purposes in the medicine of the disease of SGLT-2 suppression, described in benefit from the disease of SGLT-2 suppression selected from diabetes, diabetic keratopathy Retinopathy, diabetic neuropathy, diabetic nephropathy, wound healing delay, insulin resistance, hyperglycemia, height The rising of insulinemia, aliphatic acid blood concentration, the rising of glycerine blood concentration, hyperlipidemia, obesity, hypertriglyceridemia, X combine The diseases such as simulator sickness, atherosclerotic or hypertension.

Further aspect of the present invention provide the compound shown in Formulas I or its salt pharmaceutically accepted preparation treatment diabetes, Purposes in the medicine of the diseases such as diabetic complication, obesity, post-prandial hyperglycemia.Described diabetic complication include but not It is limited to such as diabetic retinopathy, diabetic neuropathy, diabetic nephropathy etc..

Compound shown in formula I also can be treated above-mentioned disease with the combination of other antidiabetics or delay described The progress of disease or morbidity, the example of other antidiabetics includes but not limited to: biguanides (such as, melbine or benzene second Biguanides), glucosidase inhibitor (such as, acarbose, Miglitol), insulin (such as, insulin lispro, sweet essence pancreas Island element, insulin detemir, insulin aspart, moral paddy insulin), meglitinides (such as, Repaglinide), sulfonylurea (example As, Glimepiride, glibenclamide, gliclazide, chlorpropamide, Glipizide), thiazolidinediones (such as, troglitazone, Rosiglitazone, Pioglitazone), PPAR-α/γ dual agonists (such as, Mo Gelieta), GLP-1 receptor stimulating agent (such as, Chinese mugwort Fill in that peptide, Arg34Lys26-(N-EPSILON-(N-ALPHA-Palmitoyl-L-GAMMA-glutamyl))-GLP-1[7-37]) and DPP4 inhibitor (such as Xi Gelieting, BMS-477118, Egelieting).

Compared with former medicine canagliflozin, the compound shown in Formulas I has more excellent pharmacokinetic property and (internal partly declines Phase is longer), it is expected to reduce clinical dosage, thus reduces treatment cost to allow more patient be benefited.

Detailed description of the invention:

The following examples can illustrate in greater detail the present invention, but limits the present invention the most in any form.

The preparation of embodiment 1 compound 3

Step 1): [2,3,4,5-D₄] preparation of thiophene I-2:

In pressure pipe, weigh thiophene I-1 (5.0g, 59.4mmol), add deuterated water 14ml (700mmol), be cooled to 0 P it is slowly added to after DEG C₂O₅(28.4g, 200mmol), stirs 5min under room temperature, be heated to 120 DEG C and stir 24 hours.It is cooled to 0 Add water (80ml) after DEG C, use CH₂Cl₂Extract reactant liquor (3 × 60ml).MgSO₄After drying, filtrate is distilled off at ambient pressure Obtaining oily residue (5.0g) after solvent, under normal pressure, distillation obtains compound I-24.0g, yield 80%.

It is 98.5% that LCMS surveys deuterium content.

HRMS (ESI) m/z:88.0281.

Step 2): the bromo-[3,4,5-D of 2-₃] preparation of thiophene I-3

At 0 DEG C, N-bromo-succinimide NBS (2.1g, 11.8mmol) is added to Formulas I-2 compound (0.986g, AcOD (25ml) 11.2mmol) and CHCl₃In solution (25ml), at 50 DEG C, stir reactant mixture 2 hours (until TLC detects without starting material left).Being cooled to 5-10 DEG C, add water (80ml), separates organic layer, water layer is used CH again₂Cl₂Extract (3 × 50ml).Anhydrous MgSO₄After drying, filtrate obtains residue, by residue silica gel after under reduced pressure room temperature is distilled off solvent Column chromatography (EtOac/hexane 6:94) separating-purifying product I-3, yield 80%.

¹HNMR (300MHz, DMSO-d6): δ without fragrance hydrogen peak.

HRMS (ESI) m/z:164.9321.

Step 3): 4-is to fluorophenyl [3,4,5-D₃] preparation of thiophene I-4

Addition Formulas I-3 compound (13.98g, 85.8mmol) in 1000ml eggplant-shape bottle, 4-fluorobenzoic boric acid (12.0g, 85.8mmol), Pd (PPh₃)₄(0.96g, 0.85mmol), glycol dimethyl ether 150ml, Na₂CO₃(27.3g, 258mmol's) Aqueous solution 150ml, is heated to 80 DEG C of reaction 3h.Room temperature is down to by reactant mixture, pours in frozen water, CH₂Cl₂Extraction (2 × 100ml), organic layer, anhydrous MgSO are isolated₄It is dried.Steaming solvent, crude product column chromatography for separation obtains white solid I- 413.68g, yield 90.8%.

¹HNMR (300MHz, DMSO-d6): δ 7.67～7.71 (m, 2H), 7.22～7.28 (m, 2H).

HRMS (ESI) m/z:181.0437.

It is 98.5% that LCMS surveys deuterium content.

Step 4): 2-(bromo-2 methyl-benzyls of 5-)-5-(4-fluorophenyl) [3,4-D₂] preparation of thiophene I-7

(bromo-2 aminomethyl phenyls of 5-) (5-(4-fluorophenyl) [3,4-D₂] thiophene-2-base ketone I-6:500ml eggplant-shape bottle adds I-4 (13.86g, 77.8mmol), 2-methyl-5-bromo-benzoyl chloride I-5 (19.08g, 81.7mmol), CH₂Cl₂250ml dissolves, and 0 AlCl is added at DEG C₃(11.4g, 85.6mmol), is warmed to room temperature reaction overnight after 30min.Reactant mixture is poured in frozen water, CH₂Cl₂Extraction (2 × 100ml), the anhydrous MgSO of organic layer₄It is dried.Steam solvent, in mixture, add n-hexane, filter, filter Cake n-hexane is washed, and is dried to obtain faint yellow solid I-624.3g, yield 83.2%.

¹HNMR (300MHz, CDCl₃): δ 7.62～7.66 (2H, m), 7.57 (1H, s), 7.49～7.52 (1H, dd, J₁= 8.2Hz,J₂=1.8Hz), 7.09～7.18 (3H, m), 2.33 (3H, s).

¹³C-NMR (300MHz, CDCl₃): δ 188.32,164.99,161.68,153.23,142.62,140.06, 135.36,133.14,132.71,130.47,129.48,129.43,128.26,128.15,124.00,118.80,116.41, 116.12,19.15。

HRMS (ESI) m/z:375.9908.

2-(bromo-2 methyl-benzyls of 5-)-5-(4-fluorophenyl) [3,4-D₂] thiophene I-7:

Addition I-6 (10.0g, 26.5mmol) in method one: 500ml three-necked bottle, triethyl silicane (18.5ml, 106.5mmol), CH₂Cl₂50ml, acetonitrile 50ml, be down to 0 DEG C.Boron trifluoride ether solution it is slowly added dropwise in reactant mixture (12.5ml, 100.5mmol), finishes, and is warmed to room temperature reaction overnight.Ice bath downhill reaction thing adds saturated NaHCO₃Solution 50ml, CH₂Cl₂Extraction, the anhydrous MgSO of organic layer₄It is dried.Steam solvent, obtain yellow solid I-78.9g, yield 93.6%.

Method two: 100ml bottle adds I-6 (2.0g, 5.3mmol), oxolane 20ml, NaBH₄(0.2g, 5.3mmol), it is down to 0 DEG C, adds AlCl₃(0.77g, 5.8mmol), rises to 70 DEG C of reaction 5h.Pour reactant mixture into frozen water In, EA extracts, and steams solvent, obtains white solid I-7 (1.85g), yield 96%.

¹HNMR (300MHz, CDCl₃): δ 7.47 (2H, dd, J₁=5.4Hz, J₂=8.4Hz, Ph), 7.33 (1H, s, Ph), 7.29 (1H, d, J=8.1Hz, Ph), 6.99～7.04 (3H, m, Ph), 4.06 (2H, s, Ph-CH₂-thio),2.26(3H,s, Me)。

¹³C-NMR (300MHz, CDCl₃): δ 163.71,160.51,142.10,140.33,135.28,132.16, 130.74,130.69,129.89,127.20,127.10,122.64,119.65,115.87,115.58,33.78,18.98。

HRMS (ESI) m/z:362.0113.

Step 5): (3R, 4S, 5S, 6R)-2-3-((5-(4-fluorophenyl) [3 ', 4 '-D₂] thiophene-2-base) methyl)-4-first Phenyl) preparation of-6-(methylol)-2-methoxyl group tetrahydrochysene-2H-pyrans-3,4,5-triol I-9

(3R, 4S, 5R, 6R)-3,4,5-three (trimethyl silicane alkoxyl) methyl) tetrahydrochysene-2H-pyran-2-one I-8: by Portugal Grape saccharic acid lactone (30g, 168.5mmol), anhydrous tetrahydro furan (250ml) are added sequentially in 1L there-necked flask, add N-first Base morpholine (150ml, 1.35mol).Reaction bulb is transferred in-20 degree low temperature baths, by trim,ethylchlorosilane under nitrogen protection (129ml, 1.01mol) is slowly dropped in reactant liquor in constant pressure funnel, and whole dropping process continues 30 minutes.Finish, continue Reaction bulb was transferred in oil bath pan after 15 minutes by continuous low-temp reaction, opens and is heated to reactant liquor backflow.TLC detection after 2 hours Display raw material entirely without residue, basic all for the purpose of product.After room temperature is down to by reaction bulb, addition 600ml petroleum ether, 400ml water, Being sufficiently stirred for rear separatory, TLC display water layer is without obvious product, and organic layer is containing purpose product and a small amount of N-methylmorpholine.Organic layer Add 300ml water, 25g potassium dihydrogen phosphate, be sufficiently stirred for rear separatory.Organic layer still has trace N-methylmorpholine, again adds 200ml water, 15g potassium dihydrogen phosphate, be sufficiently stirred for rear separatory, and TLC shows without obvious N-methylmorpholine and other impurity.Organic layer Steaming to obtain pale yellow oily liquid body with anhydrous sodium sulfate (100g) rotation after drying, silica gel column chromatography purifies, petroleum ether-ethyl acetate (20:1, V/V), obtains colourless oil liquid I-8 (65.5g, 140.2mmol), yield 83.2%.

(3R, 4S, 5S, 6R)-2-3-((5-(4-fluorophenyl) [3 ', 4 '-D₂] thiophene-2-base) methyl)-4-tolyl)- 6-(methylol)-2-methoxyl group tetrahydrochysene-2H-pyrans-3, in 4,5-triol I-9:100ml three-necked bottles add I-7 (1.85g, 5mmol), THF (Non-aqueous processing) 15ml, toluene (Non-aqueous processing) 15ml, N₂Under protection, at-78 DEG C, it is slowly added dropwise n-BuLi Hexane solution (2.13ml, 5.4mmol), react 20min.In reactant mixture, dropping is dissolved in 5ml toluene (anhydrous place Reason) Formulas I-8 compound (2.12g, 4.5mmol), 5min adds.Reaction 30min, adds the methyl being dissolved in 10ml absolute methanol Sulfonic acid solutions (1.29g, 13.5mmol), 5min adds.It is warmed to room temperature reaction overnight.Ice bath downhill reaction thing add saturated NaHCO₃Solution 20ml, ethyl acetate extracts, the anhydrous MgSO of organic layer₄It is dried.Steaming solvent, it is solid that column chromatography for separation obtains white Body I-9 (0.8g), yield 38%.

¹H-NMR (300MHz, DMSO-d6): δ 7.55～7.60 (2H, m, Ph), 7.43 (1H, s, Ph), 7.33 (1H, d, J =7.9Hz, Ph), 7.13～7.22 (3H, m, Ph), 4.93 (1H, d, J=5.3Hz, OH), 4.68 (1H, d, J=4.9Hz, OH), 4.63 (1H, d, J=7.3Hz, OH), 4.50 (1H, t, J=5.8Hz, OH), 4.11～4.16 (2H, dd, J₁= 15.7Hz,J₂=29.2Hz, Ph-CH₂-thio), 3.73～3.79 (1H, m, sugar), 3.51～3.64 (2H, m, sugar), 3.37～3.41 (1H, m, sugar), 3.22～3.26 (1H, m, sugar), 2.96 (3H, s, OMe), 2.90～2.92 (1H, m, sugar),2.26(3H,s,Me)。

¹³C-NMR (300MHz, DMSO-d6): δ 162.94,159.70,143.69,140.05,137.22,136.90, 135.09,130.54,129.37,128.74,126.94,126.83,126.04,115.97,115.68,100.83,77.03, 74.33,73.80,70.21,60.94,48.18,33.59,18.66。

HRMS (ESI) m/z:476.1638.

Step 6): (2S, 3R, 4R, 5S, 6R)-2-3-((5-(4-fluorophenyl) [3 ', 4 '-D₂] thiophene-2-base) methyl)- 4-tolyl) preparation of-6-(methylol)-tetrahydrochysene-2H-pyrans-3,4,5-triol 3

Addition I-9 (0.5g, 1.05mmol) in 25ml three-necked bottle, triethyl silicane (0.72ml, 4.18mmol), 8mlCHCl₂, N₂Protection, is down to-70 DEG C.Be slowly added dropwise in reactant mixture boron trifluoride ether solution (0.5ml, 3.99mmol).Rise to 0 DEG C of reaction 2h.Saturated NaHCO is added in reactant mixture₃Solution 10ml, ethyl acetate extracts, has The anhydrous MgSO of machine layer₄It is dried.Steaming solvent, column chromatography for separation obtains white solid 3 (0.3g), yield 65%.

¹H-NMR (300MHz, DMSO-d6): δ 7.56～7.60 (2H, dd, J₁=8.6Hz, J₂=5.5Hz, Ph), 7.10 ～7.25 (5H, m, Ph), 4.83 (2H, s, OH), 4.65 (1H, s, OH), 4.35 (1H, s, OH), 4.12 (2H, s, Ph-CH₂- Thio), 3.98 (1H, d, J=9.2Hz, sugar), 3.69～3.73 (1H, m, sugar), 3.47 (1H, m, sugar), 3.18 ～3.28 (4H, m, sugar), 2.26 (3H, s, Me).

¹³C-NMR (300MHz, DMSO-d6): δ 162.92,159.68,143.47,140.16,138.15,137.26, 134.83,130.49,129.56,128.98,126.93,126.82,126.15,123.32,115.91,115.62,81.24, 81.10,78.44,74.62,70.41,61.41,33.33,18.70。

HRMS (ESI) m/z:446.1536.

The preparation of embodiment 2 compound 1

Step 1): (5-bromo-2-aminomethyl phenyl) (preparation of 5-(4-fluorophenyl) thiophene-2-base ketone II-4

The bromo-5-of 2-(4-fluorophenyl) thiophene II-2: compound II-1 (4.0g, 22.47mmol) is joined 100ml single port In flask, add dichloromethane (40ml), open magnetic agitation, under condition of ice bath add N-bromosuccinimide (4.0g, 22.47mmol).It is stirred overnight under room temperature condition after continuing stirring 20 minutes.TLC shows without obvious starting material left and impurity. Reactant liquor reduced pressure concentration removes major part methylene chloride, is subsequently adding 80ml petroleum ether and is sufficiently stirred for 20 minutes, and decompression is taken out Filter, filtrate obtains about 6.9g white solid (containing part succinimide) after concentrating.Solid rejoins in 50ml petroleum ether and fills Reduce pressure after dividing stirring 20 minutes suction filtration again, is dried and obtains 5.70g snow-white solid II-2, yield after filtrate reduced in volume 95.3%.

¹H-NMR(CDCl₃, 300MHz): δ 7.44-7.48 (m, 2H), 7.00-7.09 (m, 3H), 6.95-6.97 (m, 1H).

¹³C-NMR(CDCl₃, 300MHz): δ 164.14,160.85,144.75,130.84,127.40,127.30, 123.24,116.12,115.83,111.33.

5-bromo-N-methoxyl group-N, 2-dimethylbenzamide II-3: by 2-methyl-5-bromobenzoic acid (2.15g, 10mmol) Join in 50ml single port flask, be sequentially added into dichloromethane (25ml), N, O-dimethyl hydroxylamine hydrochloride (1.07g, 11mmol), dicyclohexylcarbodiimide (2.26g, 11mmol), I-hydroxybenzotriazole one water thing (1.48g, 11mmol).Nitrogen Triethylamine (1.67ml, 12mmol) is added under gas shielded.Stirred overnight at room temperature, TLC display raw material 2-methyl-5-bromobenzoic acid is still Have relatively multiresidue, add N, O-dimethyl hydroxylamine hydrochloride (1.07g, 11mmol), dicyclohexylcarbodiimide (1.50g, 7.3mmol), triethylamine (1.0ml, 7.2mmol).After being stirred at room temperature 5 hours, TLC display raw material is without substantially residue.Reactant liquor subtracts Pressure suction filtration, filter cake dichloromethane washes twice (5ml × 2).Filtrate is sequentially added into 20ml dichloromethane and 20ml water, after separatory Water layer adds the extraction of 10ml dichloromethane.Merging organic layer, saturated aqueous common salt washs, and anhydrous sodium sulfate reduces pressure after drying and distills Product crude product.Crude product silica gel column chromatography is purified, and petroleum ether-ethyl acetate (20:1, V/V) elutes, and obtains light yellow viscous liquid II-3 (1.42g, 5.48mmol), yield 54.8%.

¹H-NMR(CDCl₃, 300MHz): δ 7.39-7.41 (m, 2H), 7.06-7.09 (m, 1H), 3.51 (s, 3H), 3.30 (s, 3H), 2.27 (s, 3H).

¹³C-NMR(CDCl₃, 300MHz): δ 136.99,134.82,133.88,132.05,131.77,128.95, 118.78,61.14,26.85,18.58.

(bromo-2 aminomethyl phenyls of 5-) (5-(4-fluorophenyl) thiophene-2-base ketone II-4: by compound II-2 (0.386g, 1.5mmol) join in dry 15ml there-necked flask, add the magnesium chips (0.096g, 6.0mmol) that watery hydrochloric acid activated, nitrogen Air in displacement reaction bottle.Absolute ether (3ml) syringe is injected in reaction bulb, opens and be heated to solvent refluxing.Will be few Being permitted iodine grain (5) to be added rapidly in reactant liquor, reactant liquor is dark-brown, continuation heating stir about reactant liquor color after 5 minutes Gradually become shallower as to colourless.Continue reaction at reflux and be directly used in grignard addition reaction after 15 minutes.By compound II-3 (0.39g, 1.5mmol) is dissolved in anhydrous tetrahydro furan (2ml), air in nitrogen displacement reaction bottle.Lattice will be made by oneself under condition of ice bath Family name's reagent (3ml, 0.5M in Ether) is slowly dropped in reaction bulb.Ice bath stirring at normal temperature is removed, after 2 hours after 5 minutes TLC shows a large amount of product and generates (standard sample comparison), is joined by reactant liquor in frozen water (6g), stands and divide after being sufficiently stirred for Liquid, water layer dichloromethane extracts (2 × 15ml).Merge organic layer, saturated aqueous common salt washing (3 × 10ml), anhydrous sodium sulfate Pale solid is steamed to obtain in dried rotation.Above-mentioned solid joins making beating stirring 30 minutes in 50ml petroleum ether, and reduce pressure suction filtration, filter Cake obtains off-white color solid II-4 (0.402g) after drying.

¹H-NMR(CDCl₃, 300MHz): δ 7.62-7.66 (m, 2H), 7.57 (d, J=1.5Hz, 1H), 7.49-7.52 (dd, J₁=1.9Hz, J₂=5.2Hz, 1H), 7.37 (d, J=3.9Hz, 1H), 7.25 (d, J=3.8Hz, 1H), 7.09-7.18 (m, 3H), 2.33 (s, 3H).

¹³C-NMR(CDCl₃, 300MHz): δ 118.29,164.99,153.27,140.08,136.68,135.33, 133.12,132.07,130.46,128.25,128.14,124.09,118.80,116.39,116.10,19.13.

Step 2): 2-(the bromo-2 methyl [1,1-D of 5-₂] benzyl) preparation of-5-(4-fluorophenyl) thiophene II-5:

Compound II-4 (2.06g, 5.51mmol) is joined in 50ml there-necked flask, adds acetonitrile (20ml), nitrogen Air in displacement reaction bottle, rapidly join deuterated sodium borohydride (0.56g, 13.78mmol), alchlor (1.10g, 8.26mmol), being transferred to by reaction bulb in 55 degree of oil bath pans, have a large amount of bubble to escape, after 10 minutes, TLC display raw material reacts Completely.Reaction bulb is cooled to room temperature, is slowly added into by reactant liquor in 30ml frozen water mixed liquor under stirring, and a large amount of whites are solid Body separates out, and reduce pressure suction filtration, and filter cake 10ml water washes twice.After filtration cakes torrefaction off-white color solid II-5 (1.64g, 4.52mmol, yield 82.0%).

¹HNMR (300MHz, CDCl₃): δ 7.45～7.49 (2H, m, Ph), 7.24～7.28 (1H, m, thio), 7.30～ 7.34 (1H, m, Ph), 6.99～7.05 (4H, m, Ph), 6.66～6.67 (1H, d, J=3.3Hz, thio), 2.26 (3H, s, Me)。

¹³C-NMR(CDCl₃, 300MHz): δ 163.79,160.52,142.12,141.86,140.24,135.26, 132.15,132.02,129.89,127.21,127.11,126.22,122.72,119.65,115.85,115.56,18.94, 15.07。

HRMS (ESI) m/z:362.0111.

Step 3): (3R, 4S, 5S, 6R)-2-3-((5-(4-fluorophenyl) thiophene-2-base) [1,1-D₂] methyl)-4-toluene Base)-6-(methylol)-2-methoxyl group tetrahydrochysene-2H-pyrans-3, the preparation of 4,5-triol II-6

Compound II-5 (1.41g, 3.88mmol) is joined 100ml be dried in there-necked flask, nitrogen displacement reaction bottle Middle air.Successively dry toluene (12ml), anhydrous tetrahydro furan (18ml) are injected reaction bulb with syringe under nitrogen protection.Will Reaction bulb is placed in the bath of-78 DEG C of low temperature, and in question response liquid, temperature drop is to after-78 DEG C, by n-butyllithium solution (1.71ml, 4.27mmol, 2.5M) it is slowly injected in reactant liquor, reactant liquor quickly becomes navy blue, and whole dropping process continues 3 minutes, instead In answering liquid, temperature is less than-66 DEG C.After 30 minutes, by be dissolved in 6ml dry toluene TMS-glucolactone (1.81g, During 3.88mmol) solution is slowly injected into reactant liquor, whole dropping process continues 3 minutes, and in keeping reactant liquor, temperature is less than-66 DEG C. Reactant liquor continues to stir 2 hours in-78 DEG C of low temperature baths.Absolute methanol by pyrovinic acid (1.49g, 15.52mmol) (12ml), during solution is slowly injected into reaction bulb, in keeping reactant liquor, temperature is less than-60 DEG C.Drip complete, reaction bulb is placed in outdoor Under the conditions of continue stirring reaction 3 hours.Under condition of ice bath, 80ml saturated sodium bicarbonate solution is slowly added in reactant liquor, fills Dividing separatory after stirring, water layer 80ml ethyl acetate is extracted twice, and merges organic layer, and saturated nacl aqueous solution (3 × 15ml) is washed Washing, anhydrous sodium sulfate revolves after drying and steams to obtain off-white color solid.Crude product silica gel column chromatography is purified, and acetate-methanol elutes (10:1, V/V), obtains white solid II-6 (1.40g, 2.94mmol, yield 75.8%).

¹H-NMR (300MHz, DMSO-d6): δ 7.55～7.60 (2H, m, Ph), 7.44 (1H, s, Ph), 7.33～7.35 (1H, d, J=7.8Hz, Ph), 7.26～7.27 (1H, d, J=3.3Hz, thio), 7.13～7.22 (3H, m, Ph), 6.76～ 6.77 (1H, d, J=3.4Hz, thio), 4.95 (1H, d, 5.3Hz, OH), 4.71 (1H, d, 4.9Hz, OH), 4.65 (1H, d, 7.2Hz, OH), 4.53 (1H, t, 5.8Hz, OH), 3.75～3.80 (1H, m, sugar), 3.52～3.65 (2H, m, sugar), 3.39～3.44 (1H, m, sugar), 3.20～3.28 (1H, m, sugar), 2.99 (3H, s, OMe), 2.26 (3H, s, Me).

¹³C-NMR (300MHz, DMSO-d6): δ 162.94,159.70,143.68,140.13,137.20,136.81, 135.05,130.53,129.34,128.70,126.94,126.83,126.12,126.01,123.33,115.95,115.66, 100.81,77.03,74.32,73.78,70.22,60.94,48.16,18.63。

HRMS (ESI) m/z:476.1638.

Step 4): (2S, 3R, 4R, 5S, 6R)-2-3-((5-(4-fluorophenyl) thiophene-2-base) [1,1-D₂] methyl)-4- Tolyl) preparation of-6-(methylol)-tetrahydrochysene-2H-pyrans-3,4,5-triol 1

Compound II-6 (1.20g, 2.52mmol) is joined in tri-mouthfuls of reaction bulbs of 50ml, be sequentially added into dichloromethane (20ml), triethyl silicane (1.20ml, 7.56mmol).Reaction bulb is placed in-50 DEG C of low temperature baths.By BFEE (0.96ml, 7.56mmol) solution is slowly injected in reaction bulb.Treat that boron trifluoride ether solution finishes, reaction bulb is placed in room temperature Under the conditions of be stirred overnight.Being slowly added in reactant liquor by 30ml saturated sodium bicarbonate solution, be sufficiently stirred for rear separatory, water layer is used 30ml ethyl acetate extracts, and merges organic layer, and anhydrous sodium sulfate revolves steaming after drying and obtains off-white color solid.Crude product silica gel column chromatography Purify, acetate-methanol wash-out (10:1, V/V), obtain white solid 1 (900mg).

¹H-NMR (300MHz, DMSO-d6): δ 7.59 (2H, dd, J₁=5.4Hz, J₂=8.7Hz, Ph), 7.27 (1H, d, J =3.5Hz, thio), 7.13～7.22 (5H, m), 6.80 (1H, d, J=3.5Hz, thio), 4.90 (2H, d, J=4.4Hz, OH), 4.71 (1H, d, J=5.5Hz, OH), 4.42 (1H, t, J=5.7Hz, OH), 3.97 (1H, d, J=9.2Hz, sugar), 3.67～3.73 (1H, m, sugar), 3.40～3.48 (1H, m, sugar), 3.12～3.27 (4H, m, sugar), 2.26 (s, 3H)。

¹³C-NMR (300MHz, DMSO-d6): δ 162.94,159.70,143.49,140.18,138.16,137.22, 134.85,130.51,129.57,128.98,126.96,126.85,126.27,126.17,123.32,115.94,115.65, 81.26,81.13,78.45,74.63,70.42,61.42,18.71。

HRMS (ESI) m/z:446.1536.

The preparation of embodiment 3 compound 4

Step 1): 2-[D₃] preparation of methyl 5-bromobenzoic acid III-2:

Weighing 2-methyl 5-bromobenzoic acid III-1 (5.0g, 23.3mmol) in microwave pressure-resistant pipe, NaOD (40%, 28.0mmol), add deuterated water 24ml (24g, 1.2mol), stir 30 minutes under room temperature, be heated to 160 DEG C and stir 4 hours.Remove Remove heavy water, add deuterated water 24ml.It is heated to 160 DEG C stir 2 hours.Add DCl after being cooled to 0 DEG C and adjust about pH=3, use CH₂Cl₂Extract reactant liquor (3 × 30ml).MgSO₄After drying, filtrate obtains oily residue after solvent is under reduced pressure distilled off III-2 (4.6g, yield 90%).

¹H NMR(CDCl₃): δ 7.70 (m, 2H), 7.25 (m, 2H).

HRMS (ESI) m/z:181.0439.

Step 2): (5-bromo-2-[D₃] aminomethyl phenyl) (the preparation of 5-(4-fluorophenyl) thiophene-2-base ketone III-3

Method one

With reference to embodiment 1 step 4) prepare compound III-3.

¹HNMR (300MHz, CDCl₃): δ 7.62～7.66 (2H, m), 7.57 (1H, m), 7.49～7.52 (1H, dd, J₁= 8.2Hz, J₂=1.8Hz), 7.36～7.38 (1H, d, J=3.8Hz), 7.24～7.25 (1H, d, J=3.8Hz), 7.09～ 7.18 (3H, m).

HRMS (ESI) m/z:376.9961.

Method two

In microwave pressure-resistant pipe, add compound II-4 (5.0g, 13.3mmol), NaOD (3.0mmol), add deuterated water 24ml (24g, 1.2mol), is heated to 160 DEG C and stirs 4 hours.Remove heavy water, add fresh deuterated water 12ml and THF (12ml).It is heated to 160 DEG C stir 4 hours.It is cooled to room temperature, uses CH₂Cl₂Extract reactant liquor (3 × 30ml).MgSO₄It is dried After, filtrate obtains oily residue III-3 (4.3g, yield 84%) after solvent is under reduced pressure distilled off.

Step 3): 2-(5-bromo-2-[D₃] methyl-benzyl) preparation of-5-(4-fluorophenyl) thiophene III-4

Joining in there-necked flask by compound III-3 (2.0g, 5.31mmol), add THF (20ml), nitrogen displacement is anti- Answer air in bottle, rapidly join aluminium lithium hydride (0.50g, 13.3mmol), reaction bulb is transferred in 55 degree of oil bath pans, have a large amount of Bubble escapes, and after 10 minutes, TLC display raw material has reacted complete.Reaction bulb is cooled under 0 DEG C, stirring join 0.5ml Frozen water, 0.5ml 15%NaOH and 1.5ml frozen water are in reaction mixture, and after stirring 2 hours, a large amount of white solids separate out, and subtract Pressure suction filtration, filter cake 10ml THF washes twice.Filtrate is distilled off THF after drying, obtain off-white color solid III-4 (1.64g, 4.52mmol, yield 82.0%).

¹HNMR (300MHz, CDCl₃): δ 7.45～7.49 (2H, m), 7.24～7.34 (2H, m), 6.99～7.05 (4H, M), 6.66～6.67 (1H, d, J=3.3Hz), 4.04 (2H, s).

HRMS (ESI) m/z:363.0112.

Step 4): (3R, 4S, 5S, 6R)-2-3-((5-(4-fluorophenyl) thiophene-2-base) methyl)-4-[D₃] tolyl)- The preparation of 6-methylol-2-methoxyl group tetrahydrochysene-2H-pyrans-3,4,5-triol III-5

With reference to embodiment 1 step 5) prepare compound III-5.

¹HNMR (300MHz, DMSO-d6): δ 7.55～7.60 (2H, m, Ph), 7.42 (1H, s, Ph), 7.32～7.34 (1H, d, J=7.0Hz, Ph), 7.26～7.27 (1H, d, J=3.4Hz, thio), 7.13～7.22 (3H, m, Ph), 6.76～ 6.77 (1H, d, J=3.3Hz, thio), 4.92 (1H, s, OH), 4.50～4.64 (3H, m, OH), 4.11,4.16 (each 1H, S, Ph-CH₂-thio), 3.74～3.77 (1H, m, sugar), 3.51～3.62 (2H, m, sugar), 3.37 (1H, m, Sugar), 3.17～3.25 (1H, m, sugar), 2.96 (3H, s, OMe), 2.90 (1H, s, sugar).

HRMS (ESI) m/z:477.1705.

Step 5): (2S, 3R, 4R, 5S, 6R)-2-3-((5-(4-fluorophenyl) thiophene-2-base) methyl)-4-[D₃] toluene Base) preparation of-6-(methylol)-tetrahydrochysene-2H-pyrans-3,4,5-triol 4

With reference to embodiment 1 step 6) prepare compound 4.

¹HNMR (300MHz, DMSO-d6): δ 7.56～7.61 (2H, dd, J₁=8.6Hz, J₂=5.5Hz, Ph), 7.26～ 7.27 (1H, d, J=3.5Hz, thio), 7.10～7.22 (5H, m, Ph), 6.78～6.80 (1H, d, J=3.4Hz, thio), 4.89～4.90 (2H, d, J=4.4Hz, OH), 4.69～4.71 (1H, d, J=5.6Hz, OH), 4.39～4.43 (1H, t, J= 5.7Hz, OH), 4.11,4.12 (each 1H, s, Ph-CH₂-thio), 3.94～3.98 (1H, d, J=9.3Hz, sugar), 3.67～3.72 (1H, m, sugar), 3.40～3.47 (1H, m, sugar), 3.12～3.27 (4H, m, sugar).

HRMS (ESI) m/z:447.1591.

The preparation of embodiment 4 compound 5

Step 1): the 2-(bromo-2 [D of 5-₃] methyl [1 ', 1 '-D₂] benzyl) preparation of-5-(4-fluorophenyl) thiophene IV-1

With reference to embodiment 2 step 2) prepare compound IV-1.

¹HNMR (300MHz, CDCl₃): δ 7.44～7.47 (2H, m), 7.23～7.34 (2H, m), 6.97～7.05 (4H, M), 6.64～6.67 (1H, d, J=3.3Hz).

HRMS (ESI) m/z:365.0237.

Step 2): (3R, 4S, 5S, 6R)-2-3-((5-(4-fluorophenyl) thiophene-2-base) [1,1-D₂] methyl)-4-[1, 1,1-D₃] tolyl) preparation of-6-(methylol)-2-methoxyl group tetrahydrochysene-2H-pyrans-3,4,5-triol IV-2

With reference to embodiment 1 step 5) prepare compound IV-2.

¹HNMR (300MHz, DMSO-d6): δ 7.54～7.61 (2H, m, Ph), 7.42 (1H, s, Ph), 7.32～7.34 (1H, d, J=7.0Hz, Ph), 7.26～7.27 (1H, d, J=3.4Hz, thio), 7.13～7.22 (3H, m, Ph), 6.76～ 6.77 (1H, d, J=3.3Hz, thio), 4.92 (1H, s, OH), 4.50～4.64 (3H, m, OH), 3.74～3.77 (1H, m, Sugar), 3.51～3.62 (2H, m, sugar), 3.37 (1H, m, sugar), 3.17～3.25 (1H, m, sugar), 2.96 (3H, s, OMe), 2.90 (1H, s, sugar).

HRMS (ESI) m/z:479.1804.

Step 3): (2S, 3R, 4R, 5S, 6R)-2-3-((5-(4-fluorophenyl) thiophene-2-base) [1,1-D₂] methyl)-4- [1,1,1-D₃] tolyl) preparation of-6-(methylol)-tetrahydrochysene-2H-pyrans-3,4,5-triol 5

With reference to embodiment 1 step 6) prepare compound 5.

¹HNMR (300MHz, DMSO-d6): δ 7.55～7.60 (2H, dd, J₁=8.6Hz, J₂=5.5Hz, Ph), 7.26～ 7.27 (1H, d, J=3.5Hz, thio), 7.10～7.22 (5H, m, Ph), 6.78～6.80 (1H, d, J=3.4Hz, thio), 4.89～4.90 (2H, d, J=4.4Hz, OH), 4.69～4.71 (1H, d, J=5.6Hz, OH), 4.39～4.43 (1H, t, J= 5.7Hz, OH), 3.94～3.98 (1H, d, J=9.3Hz, sugar), 3.67～3.72 (1H, m, sugar), 3.40～3.47 (1H, m, sugar), 3.12～3.27 (4H, m, sugar).

HRMS (ESI) m/z:449.1771.

The preparation of embodiment 5 compound 13

Step 1): 2-(5-bromo-2-[D₃] methyl-benzyl)-5-(4-fluorophenyl) [3,4-D₂] thiophene V-2:

(5-bromo-2-[D₃] aminomethyl phenyl) (5-(4-fluorophenyl) [3,4-D₂] thiophene-2-base ketone V-1: with reference to embodiment 1 step Rapid 4) method prepares compound V-1.

¹HNMR (300MHz, CDCl₃): δ 7.61～7.65 (2H, m), 7.57 (1H, m), 7.49～7.52 (1H, dd, J₁= 8.2Hz, J₂=1.8Hz), 7.09～7.18 (3H, m).

HRMS (ESI) m/z:379.0040.

2-(5-bromo-2-[D₃] methyl-benzyl)-5-(4-fluorophenyl) [3,4-D₂] thiophene V-2: with reference to embodiment 3 step 3) Method prepare compound V-2.

¹HNMR (300MHz, CDCl₃): δ 7.45～7.49 (2H, m), 7.24～7.34 (2H, m), 6.99～7.05 (4H, M), 6.66～6.67 (1H, d, J=3.3Hz), 4.06 (2H, s).

HRMS (ESI) m/z:365.0247.

Step 2): (3R, 4S, 5S, 6R)-2-3-((5-(4-fluorophenyl) [3 ', 4 '-D2] thiophene-2-base) methyl)-4- [D₃] tolyl)-6-(methylol)-2-methoxyl group tetrahydrochysene-2H-pyrans-3,4,5-triol V-3

With reference to embodiment 1 step 5) method prepare compound V-3.

¹HNMR (300MHz, DMSO-d6): δ 7.55～7.60 (2H, m, Ph), 7.42 (1H, s, Ph), 7.32～7.34 (1H, d, J=7.0Hz, Ph), 7.13～7.22 (3H, m, Ph), 4.92 (1H, s, OH), 4.50～4.64 (3H, m, OH), 4.11,4.16 (each 1H, s, Ph-CH₂-thio), 3.74～3.77 (1H, m, sugar), 3.51～3.62 (2H, m, Sugar), 3.37 (1H, m, sugar), 3.17～3.25 (1H, m, sugar), 2.96 (3H, s, OMe), 2.90 (1H, s, sugar)。

¹³C-NMR (DMSO, 300MHz): δ 162.97,159.73,143.80,140.16,137.23,136.90, 135.10,130.55,129.38,128.76,126.96,126.86,126.14,126.05,123.35,115.59,115.69, 100.84,77.06,74.36,73.81,70.25,60.98,48.20,33.63.

HRMS (ESI) m/z:476.1638.

Step 3): (2S, 3R, 4R, 5S, 6R)-2-3-((5-(4-fluorophenyl) [3 ', 4 ' D₂] thiophene-2-base) methyl)-4- [1,1,1-D₃] tolyl)-6-(methylol)-tetrahydrochysene-2H-pyrans-3,4,5-triol 13.

With reference to embodiment 1 step 6) method prepare compound 13.

¹HNMR (300MHz, DMSO-d6): δ 7.56～7.61 (2H, dd, J₁=8.6Hz, J₂=5.5Hz, Ph), 7.10～ 7.22 (5H, m, Ph), 4.89～4.90 (2H, d, J=4.4Hz, OH), 4.69～4.71 (1H, d, J=5.6Hz, OH), 4.39 ～4.43 (1H, t, J=5.7Hz, OH), 4.11,4.12 (each 1H, s, Ph-CH₂-thio), 3.94～3.98 (1H, d, J= 9.3Hz, sugar), 3.67～3.72 (1H, m, sugar), 3.40～3.47 (1H, m, sugar), 3.12～3.27 (4H, m, sugar)。

HRMS (ESI) m/z:449.1701.

The preparation of embodiment 6 compound 10

Step 1): 2-(5-bromo-2-[D₃] methyl [D₂] benzyl)-5-(4-fluorophenyl) [3,4-D₂] thiophene VI-1

With reference to embodiment 2 step 2) method prepare compound VI-1.

¹HNMR (300MHz, CDCl₃): δ 7.45～7.49 (2H, m), 7.23～7.34 (2H, m), 6.97～7.05 (3H, m)。

HRMS (ESI) m/z:367.0411.

Step 2): (3R, 4S, 5S, 6R)-2-3-((5-(4-fluorophenyl) [3 ', 4 '-D₂] thiophene-2-base) [D₂] methyl)- 4-[D₃] tolyl)-6-(methylol)-2-methoxyl group tetrahydrochysene-2H-pyrans-3,4,5-triol VI-2.

With reference to embodiment 2 step 3) prepare compound VI-2.

¹HNMR (300MHz, DMSO-d6): δ 7.53～7.60 (2H, m, Ph), 7.42 (1H, s, Ph), 7.32～7.34 (1H, d, J=7.0Hz, Ph), 7.13～7.22 (3H, m, Ph), 4.92 (1H, s, OH), 4.50～4.64 (3H, m, OH), 3.74 ～3.77 (1H, m, sugar), 3.51～3.62 (2H, m, sugar), 3.37 (1H, m, sugar), 3.17～3.25 (1H, m, Sugar), 2.95 (3H, s, OMe).

HRMS (ESI) m/z:481.1912.

Step 3): (2S, 3R, 4R, 5S, 6R)-2-3-((5-(4-fluorophenyl) [3 ', 4 ' D₂] thiophene-2-base) [D₂] first Base)-4-[1,1,1-D₃] tolyl)-6-(methylol)-tetrahydrochysene-2H-pyrans-3,4,5-triol 10

With reference to implement 1 step 6) method prepare compound 10.

¹HNMR (300MHz, DMSO-d6): δ 7.56～7.61 (2H, dd, J₁=8.6Hz, J₂=5.5Hz, Ph), 7.10～ 7.22 (5H, m, Ph), 4.89～4.90 (2H, d, J=4.4Hz, OH), 4.69～4.71 (1H, d, J=5.6Hz, OH), 4.39 ～4.43 (1H, t, J=5.7Hz, OH), 3.94～3.98 (1H, d, J=9.3Hz, sugar), 3.67～3.72 (1H, m, Sugar), 3.40～3.47 (1H, m, sugar), 3.12～3.27 (4H, m, sugar).

HRMS (ESI) m/z:451.1814.

The preparation of embodiment 7 compound 7:

Step 1): 2-(5-bromo-2-methylbenzene [1 ', 1 '-D₂] benzyl 5-(4-fluorophenyl) [2 ', 3 '-D₂] thiophene VII-1

With reference to embodiment 1 step 4) method prepare compound VII-1.

¹H-NMR(CDCl₃, 300MHz): δ 7.47 (2H, dd, J₁=5.4Hz, J₂=8.4Hz, Ph), 7.34 (1H, s, Ph), 7.29 (1H, d, J=8.1Hz, Ph), 6.99-7.04 (3H, m, Ph), 2.26 (3H, s, Me).

¹³C-NMR (300MHz, DMSO-d6): δ 163.78,160.51,142.05,141.83,140.25,135.27, 132.16,132.03,130.74,130.70,129.90,127.21,127.10,119.65,115.87,115.58,18.97。

Step 2): (3R, 4S, 5S, 6R)-2-3-((5-(4-fluorophenyl) [3 ', 4 '-D₂] thiophene-2-base) [D₂] methyl)- 4-tolyl)-6-(methylol)-2-methoxyl group tetrahydrochysene-2H-pyrans-3,4,5-triol VII-2

With reference to embodiment 2 step 3) method prepare compound VII-2.

¹HNMR (300MHz, DMSO-d6): δ 7.55-7.60 (2H, m, Ph), 7.43 (1H, s, Ph), 7.33 (1H, d, J= 7.0Hz, Ph), 7.13-7.22 (3H, m, Ph), 4.88 (1H, d, J=5.3Hz, OH), 4.63 (1H, d, J=4.9Hz, OH), 4.57 (1H, d, J=7.3Hz, OH), 4.46 (1H, d, J=6.0Hz, OH), 3.73-3.79 (1H, m, sugar), 3.52-3.64 (2H,m,sugar),3.39-3.42(1H,m,sugar),3.19-3.24(1H,m,sugar),2.96(3H,s,OMe),2.90- 2.93(1H,m,sugar),2.26(3H,s,Me)；

¹³C-NMR (300MHz, DMSO-d6): δ 162.92,159.68,143.66,140.04,137.19,136.79, 135.03,130.51,129.32,128.68,126.91,126.81,125.99,123.33,115.92,115.64,100.79, 77.02,74.32,73.77,70.22,60.94,48.15,18.60。

HRMS (ESI) m/z:478.1713.

Step 3): (2S, 3R, 4R, 5S, 6R)-2-3-((5-(4-fluorophenyl) [3 ', 4 ' D₂] thiophene-2-base) [D₂] first Base)-4-tolyl)-6-(methylol)-tetrahydrochysene-2H-pyrans-3,4,5-triol 7

With reference to embodiment 1 step 6) method prepare compound 7.

¹HNMR (300MHz, DMSO-d6): δ 7.56-7.60 (m, 2H, dd, J₁=8.6Hz, J₂=5.5Hz, Ph), 7.10- 7.25 (5H, m, Ph), 4.86 (2H, d, J=4.4Hz, OH), 4.66 (1H, d, J=5.0Hz, OH), 4.37 (1H, t, J= 5.7Hz, OH), 3.98 (1H, d, J=9.1Hz, sugar), 3.69-3.74 (1H, m, sugar), 3.45-3.48 (1H, m, sugar),3.15-3.29(4H,m,sugar),2.26(s,3H)。

¹³C-NMR (300MHz, DMSO-d6): δ 162.94,159.69,143.39,140.18,138.16,137.21, 134.83,130.47,129.57,128.98,126.94,126.83,126.16,123.22,115.92,115.63,81.26, 81.11,78.46,74.64,70.43,61.43,33.33,18.70。

HRMS (ESI) m/z:448.1618.

The preparation of embodiment 8 compound 6:

(2S, 3R, 4R, 5S, 6R)-2-3-((5-(4-fluorophenyl) thiophene-2-base) [D₂] methyl)-4-tolyl)-6- (methylol)-tetrahydrochysene-2D-pyrans-3,4,5-triol 6

With reference to embodiment 1 step 6) method prepare compound 6.

¹HNMR (300MHz, DMSO-d6): δ 7.56～7.61 (2H, dd, J₁=8.6Hz, J₂=5.5Hz, Ph), 7.10～ 7.22 (5H, m, Ph), 4.89～4.90 (2H, d, J=4.4Hz), 4.69～4.71 (1H, d, J=5.6Hz), 4.39～4.43 (1H, t, J=5.7Hz), 3.67～3.72 (1H, m, sugar), 3.40～3.47 (1H, m, sugar), 3.12～3.27 (4H, M, sugar), 2.25 (s, 3H).

HRMS (ESI) m/z:448.1618.

The preparation of embodiment 9 compound 28

Rapid 1): the bromo-5-of 2-(4-fluorophenyl) [2,3-D₂] thiophene phenol IX-1

Weighing type II-1 compound (5.0g, 28.1mmol) in pressure pipe, adds deuterated water 14ml (700mmol), cold But after 0 DEG C, it is slowly added to P₂O₅(28.4g, 200mmol), stirs 5min under room temperature, be heated to 120 DEG C and stir 24 hours.Cold But after 0 DEG C, add water (80ml), use CH₂Cl₂Extract reactant liquor (3 × 60ml).MgSO₄After drying, filtrate is distilled at ambient pressure Oily residue Formulas I-4 compound (5.0g) is obtained after removing solvent.

¹H-NMR(CDCl₃, 300MHz): δ 7.71-7.67 (m, 2H), 7.22-7.28 (t, 2H).

HRMS (ESI) m/z:181.0494.

Replace Formula II-1 compound with Formulas I-4 compound, and with reference to embodiment 2 step 1) described in method formula IX-1 Compound.

¹H-NMR (CDCl3,300MHz): δ 7.51 (m, 2H), 7.02 (m, 2H).

HRMS (ESI) m/z:257.9481.

Rapid 2): (bromo-2 aminomethyl phenyls of 5-) (5-(4-fluorophenyl) [2,3-D₂] thiophene phenol-2-base ketone IX-2

Replace Formula II-2 compound with Formula IX-1 compound, and with reference to embodiment 2 step 1) described in method formula IX- 2 compounds.

¹HNMR (300MHz, CDCl3): δ 7.62～7.66 (2H, m), 7.57 (1H, s), 7.49～7.52 (1H, dd, J1 =8.2Hz, J2=1.8Hz), 7.09～7.18 (3H, m), 2.33 (3H, s).

HRMS (ESI) m/z:375.9906.

Rapid 3): (5-bromo-2-aminomethyl phenyl) (5-(-4-fluorophenyl)-[2,3-D₂] thiophene) [D]-methyl alcohol IX-3

Formula IX-2 compound (2.65g, 7.06mmol) is joined in 100mL single port flask, is sequentially added into acetonitrile (25mL)、[D₄] sodium borohydride (0.40g, 10.87mmol).Unlatching is heated to reactant liquor backflow, and after 3 hours, TLC shows raw material Reaction is completely.Reactant liquor joins in 20mL heavy water under stirring after being cooled to room temperature, has more solid to separate out.Mixed liquor subtracts Pressure suction filtration.Faint yellow solid IX-3 (2.51g) is obtained after filtration cakes torrefaction.

¹HNMR (300MHz, CDCl3): δ 7.79 (s, 1H), 7.52-7.47 (m, 2H), 7.33 (d, 1H), 7.06-6.98 (m,3H),6.66(d,1H),2.25(s,3H)。

HRMS (ESI) m/z:379..0123.

Step 4): ((5-bromo-2-aminomethyl phenyl) (5-(-4-fluorophenyl)-[2,3-D₂] thiophene) [D] methoxyl group) trimethyl Silane IX-4

Compound IX-3 (1.5g, 3.95mmol) is joined in 25mL there-necked flask, is sequentially added into oxolane (15mL), N-methylmorpholine (0.61mL, 5.13mmol).Reaction bulb is transferred in-20 DEG C of low-temp reaction baths, question response liquid temperature Degree is slowly added to trim,ethylchlorosilane (0.61mL, 4.74mmol) after being down to-20 DEG C.React after continuing stirring under low temperature 5 minutes Bottle is transferred to be stirred at room temperature, and after three hours, TLC display starting material left is less.30mL petroleum ether, 20mL water are added sequentially to instead Answering in liquid, be sufficiently stirred for rear separatory, aqueous phase 20mL petroleum ether extracts once again, separatory, merges organic phase, organic phase 15mL Saturated potassium dihydrogen phosphate washs, and anhydrous sodium sulfate revolves after drying and steams to obtain light yellow viscous liquid.Crude product column chromatography purifies, stone Oil ether-ethyl acetate (10:1, V/V) system elutions, (1.5g, 3.32mmol receive to obtain light yellow viscous liquid Formula IX-4 compound Rate 84.1%).

¹HNMR (300MHz, CDCl3): δ 7.79 (s, 1H), 7.52-7.47 (m, 2H), 7.32 (d, 1H), 7.06-6.98 (m,2H),2.25,(s,3H),0.14(s,9H)。

HRMS (ESI) m/z:451.0514.

Step 5): (2S, 3R, 4R, 5S, 6R)-2-3-((5-(4-fluorophenyl) [2,3-D₂] thiophene-2-base) hydroxyl [D] first Base)-4-tolyl)-6-(methylol)-2-methoxyl group tetrahydrochysene-2H-pyrans-3,4,5-triol IX-5

Compound IX-4 (1.61g, 3.55mmol) is joined 100mL be dried in there-necked flask, nitrogen displacement reaction bottle Middle air.Successively dry toluene (10mL), anhydrous tetrahydro furan (15mL) are injected reaction bulb with syringe under nitrogen protection.Will Reaction bulb is placed in the bath of-78 DEG C of low temperature, and in question response liquid, temperature drop is to after-78 DEG C, by n-butyllithium solution (1.56mL, 3.9mmol, 2.5M) it is slowly injected in reactant liquor, reactant liquor quickly becomes navy blue, and whole dropping process continues 10 minutes, protects In holding reactant liquor, temperature is less than-65 DEG C.After 30 minutes, by be dissolved in 50mL dry toluene TMS-glucolactone I-8 (1.66g, During 3.55mmol) solution is slowly injected into reactant liquor, whole dropping process continues 10 minutes, and in keeping reactant liquor, temperature is less than-61 DEG C. Reactant liquor continues to stir 2 hours in-78 DEG C of low temperature baths.By the absolute methanol (15mL) of pyrovinic acid (1.36g, 14.2mmol) Solution is slowly injected in reaction bulb, and in keeping reactant liquor, temperature is less than-60 DEG C.Drip complete, reaction bulb is placed under outdoor conditions Continue stirring reaction 3 hours.Under condition of ice bath, 30mL saturated sodium bicarbonate solution is slowly added in reactant liquor, is sufficiently stirred for Rear separatory, water layer 20mL ethyl acetate is extracted twice, and merges organic layer, and saturated nacl aqueous solution (3 × 10mL) washs, anhydrous Sodium sulphate revolves after drying and steams to obtain off-white color solid.Crude product silica gel column chromatography is purified, acetate-methanol wash-out (10:1, V/ V), white solid Formula IX-5 compound (1.12g, yield 63.9%) is obtained.

¹HNMR (300MHz, CDCl₃): δ 1H-NMR (300MHz, DMSO-d6): δ 7.57～7.65 (3H, m, Ph), 7.36 ～7.38 (1H, m, thio), 7.11～7.29 (m, 3H, Ph), 4.93 (1H, d, 5.3Hz, OH), 4.64～4.69 (2H, m, OH), 4.49～4.51 (1H, m, OH), 3.73～3.76 (1H, m, sugar), 3.54～3.61 (2H, m, sugar), 3.38～ 3.43 (1H, m, sugar), 3.20～3.30 (1H, m, sugar), 2.99 (3H, s, OMe), 2.25 (3H, s, Me).

HRMS (ESI) m/z:493.1652.

Step 6): (2S, 3R, 4R, 5S, 6R)-2-3-((5-(4-fluorophenyl) [3 ', 4 '-D₂] thiophene-2-base) methyl)- 4-tolyl)-6-(methylol)-tetrahydrochysene-2H-pyrans-3,4,5-triol 28

Addition Formula IX-5 compound (0.5g, 1.03mmol) in 25ml three-necked bottle, triethyl silicane (0.72ml, 4.18mmol), 8ml CHCl₂, N₂Protection, is down to-70 DEG C.Boron trifluoride ether solution it is slowly added dropwise in reactant mixture (0.5ml, 3.99mmol).Rise to 0 DEG C of reaction 2h.Saturated NaHCO is added in reactant mixture₃Solution 10ml, ethyl acetate Extraction, the anhydrous MgSO of organic layer₄It is dried.Steaming solvent, column chromatography for separation obtains white solid formula 28 compound (0.3g), yield 65%.

¹H-NMR (300MHz, DMSO-d6): δ 7.56～7.60 (2H, dd, J1=8.6Hz, J2=5.5Hz, Ph), 7.10 ～7.25 (5H, m, Ph), 4.83 (2H, s, OH), 4.65 (1H, s, OH), 4.35 (1H, s, OH), 4.12 (1H, s, Ph-CHD- Thio), 3.98 (1H, d, J=9.2Hz, sugar), 3.69～3.73 (1H, m, sugar), 3.47 (1H, m, sugar), 3.18 ～3.28 (4H, m, sugar), 2.26 (3H, s, Me).

¹³C-NMR (300MHz, DMSO-d6): δ 162.92,159.68,143.47,140.16,138.15,137.26, 134.83,130.49,129.56,128.98,126.93,126.82,126.15,123.32,115.91,115.62,81.24, 81.10,78.44,74.62,70.41,61.41,33.33(m),18.70。

HRMS (ESI) m/z:447.1593.

Normal SD rats sugar tolerance and glucose in urine are drained by embodiment 9 canagliflozin, compound 1, compound 3 and compound 7 The impact of amount

One) test method

Rat [CD (SD) body weight 160-180 gram] 20, fasting 18h can't help water, is randomly divided into 5 groups by blood sugar and body weight: Normal group, canagliflozin (1.0mg/kg) group, compound 1 (1.0mg/kg) group, compound 3 (1.0mg/kg) group, chemical combination Thing 7 (1.0mg/kg) group, gavage gives relative medicine (1.0mg/kg), and normal group gives equal-volume water for injection.After administration 30min, ig give rat glucose 3g/kg, and administration volume is 1.5ml/100g, and Rat Fast terminates to experiment, and tail point is taken a blood sample Measure before being administered and to 0.5h after GLU, 1,2,4,6,8, the blood sugar of 24h, collect and give after glucose 02,24,4-6,6-10, 10 24h rat urine, (4,6h collect every rat after urine and supplement water, ig 5mL water for injection), measure urine volume and take After 1ml, 1000r/min*5min are centrifugal, measure glucose in urine.Testing index and computational methods are as follows:

(1) AUC (0～t_nBefore)=(administration+be administered after t₀)*t₀/ 2+ (t after administration₀T after+administration₁)*(t₁-t₀)/2+ (gives T after medicine₁T after+administration₂)*(t₂-t₁)/2+......+ (t after administration_n-1T after+administration_n)*(t_n-t_n-1)/2

(2) blood sugar rate of descent (%)=(normal group-administration group)/normal group × 100%

(3) blood sugar AUC rate of descent (%)=(normal group blood sugar AUC-administration group blood sugar AUC)/normal group blood sugar AUC × 100%

(4) sugar excretion=glucose in urine * volume of urine

(5) sugar excretion rate=sugar excretion/give sugar measure

Two) data statistics processing method

Data press statistical requirements, represent by mean+SD (x ± s), and all data use Microsoft Excel Software carries out t inspection, and analysis result.

Three) experimental result

(1) impact on normal rat sugar tolerance

After rats in normal control group gavage gives glucose, 30min blood glucose value significantly raises, and 60min blood sugar reaches the highest, Declining occurs in 120min blood glucose value.Compare with Normal group, except compound 1 treated animal is under rat blood sugar time graph after sugar Area AUC (0-24) conspicuousness reduces beyond (p < 0.05), each group rat glucose load 0.5h, 1,2,4,6,8,24h blood sugar with Different periods blood sugar AUC is without significant difference.To 2h after sugar rat blood sugar rate of descent is respectively as follows: 15% ,-13% ,-8% ,- 4%, rat blood sugar AUC (0-2h) rate of descent is respectively as follows: 5%, 3%, 0%, 0%；To 24h after sugar, rat blood sugar rate of descent is divided Not being: 15%, 18%, 6%, 13%, rat blood sugar AUC (0-24h) rate of descent is respectively as follows: 13%, 12%, 4%, 7%.Result It is shown in Table 1-4.

Table 1. on the impact of glucose load rat blood sugar (N=4)

Table 2. on the impact of glucose load rat blood sugar-AUC (N=4)

The impact that glucose load rat blood sugar AUC is declined by table 3.

The impact that glucose load rat blood sugar is declined by table 4.

(2) impact on normal rat urine sugar excretion

Compare with normal group, each group of glucose excretion that all can increase in glucose load rat 24h, descending be followed successively by Compound 1 (1mg/kg) group ≈ canagliflozin (1mg/kg) group ＞ compound 7 (1mg/kg) group ≈ compound 3 (1mg/kg) group. Canagliflozin, compound 1, compound 3, compound 7 (1mg/kg) group rat 0 10h glucose in urine excretion rate is respectively as follows: 11.6%, 14.7%, 10.5%, 11.2%, difference is extremely notable (P ＜ 0.01) compared with Normal group；Each group rat 10-24h glucose in urine row The rate of letting out is respectively as follows: 2.4%, 1.5%, 2.2%, 1.6%.Show arrange sugar be concentrated mainly on 0～10h, 10～24h periods arrange sugar measure Less.Each experimental group rat is more or less the same in row's sugar amount of 0-2,2-4,4-6,6-10,10-24h, and urine glucose lowering effect is more slow With, and corresponding to its hypoglycemic activity trend.The results are shown in Table 5-10.

Glucose load rat different time sections urine row's sugar is measured by table 5. to be affected (N=4)

Note: compared with Normal group, * P < 0.05, * * P < 0.01

Table 6. on glucose load rat different time sections urine volume affect (N=4)

Note: compared with Normal group, * P < 0.05, * * P < 0.01.

Table 7. on the impact of glucose load rat different time sections glucose in urine excretion rate (N=4)

Note: compared with Normal group, * P < 0.05, * * P < 0.01.

Table 8. on glucose load rat accumulation urine row sugar amount impact (N=4)

Note: compared with Normal group, * P < 0.05, * * P < 0.01.

Table 9. on glucose load rat accumulation urine volume affect (N=4)

Note: compared with Normal group, * P < 0.05, * * P < 0.01.

Table 10. on the impact of glucose load rat accumulation glucose in urine excretion rate (N=4)

Note: compared with Normal group, * P < 0.05, * * P < 0.01.

Four) brief summary

It is followed successively by normal SD rats hypoglycemic activity is descending: compound 1 (1mg/kg) group ≈ canagliflozin (1mg/ Kg) group ＞ compound 7 (1mg/kg) group ≈ compound 3 (1mg/kg) group.

Descending to the Excretion of glucose in urine it is: compound 1 (1mg/kg) group ＞ canagliflozin (1mg/kg) group ＞ization Compound 7 (1mg/kg) group ≈ compound 3 (1mg/kg) group.

Embodiment 10 canagliflozin and compound 1 metabolic stability Journal of Sex Research in people's hepatomicrosome

(1) external temperature incubates operation

1) taking out people's hepatomicrosome 3 to manage (often pipe 50 μ L, 20mg/mL), often pipe liver particle body temperature incubates solution system: 395 μ LPBS, adds 50 μ L hepatomicrosome solution (5.0mg/mL), 5 μ L substrates, mixes and get final product.

2) weigh about 8.4mg NADPH and add 500 μ L PBS dissolvings, add 500 μ L MgCl₂Mixing.

3) divide and take 450 μ L (n=2) temperature and incubate solution constant-temperature table temperature and incubate, incubate 5min in 37 DEG C of pre-temperature.NADPH mixing is molten Liquid is (containing MgCl₂) also to incubate 5min in 37 DEG C of pre-temperature stand-by.

4) drawing 54 μ L temperature and incubate solution in 1.5mL Eppendorf pipe, add 6 μ L pH7.4PBS buffer solutions, vortex mixes Even, it is 0min sample.

5) residue temperature is incubated in solution every part and is added 44 μ L NADPH mixed solutions (containing MgCl₂) start and react (the most organic The ratio of solvent is 1%), 37 DEG C of temperature are incubated and timing.After arriving the setting time, shift 60 μ L sample to adding 120 μ L reactions The 1.5mL Eppendorf pipe of stop buffer terminates reaction.

6) vortex mixing 3min, in 4 DEG C, 12000rpm be centrifuged 3min, took for 80 μ L of supernatant nights, add 80 μ L pure water, mixing, Carry out LC-MS/MS analysis.

7) if any anomaly and operation, timely record.

(2) microsome sample treatment

Draw 60 μ L microsome samples, add the ice acetonitrile precipitation of 200 μ L containing the internal standard, after vortex concussion 5min, centrifugal (13800rpm, 4 DEG C) 10min.TQ1501-754a, TQ1501-001a, DEX Aspirate supernatant 80 μ L, adds in 96 orifice plates The acetonitrile of the 1:1 of 80 μ L: water, vortex mixes, and 1 μ L sample introduction is analyzed.

(3) data acquisition and processing (DAP) method

This time test mainly sees compound elimination situation in vitro, using 0 as reference, with the ratio with internal standard peak area Value compares as data.The most do not do corresponding calibration curve and the Quality Control of each compound.

(4) estimation of pharmacokinetic parameters

Clearance rate computing formula:V/M=1/protein concentration

(5) measured data result

I. the right Mei Shafen of positive control (DEX) result

External 1 μM of DEX temperature incubates 120min, sees table 11 relative to 0min change numerical value.

The external 1 μM of DEX temperature of table 11 is incubated relative 0min and is remained percentage

The most external canagliflozin and compound 1 temperature incubate result

Canagliflozin, compound 1 temperature are incubated 90min, sees table 12 relative to 0min change numerical value.

Table 12 canagliflozin, compound 1 temperature are incubated in 90min, remain percentage relative to 0min

The most external temperature incubates parametric fitting results

Positive drug, canagliflozin, compound 1 parametric fitting results are shown in Table 13.

Table 13 positive drug, canagliflozin, compound 1 parametric fitting results

(6) brief summary

1) half-life of positive drug dextromethorphan is 34.79min, between 30～50min, meets the requirements, prompting experiment Reliable results.

2) canagliflozin and compound 1 have certain metabolism in people's hepatomicrosome in 90min, but the half-life is equal > 200min, Prompting metabolism is slowly.

3) compound 1 of same concentrations is more longer than canagliflozin half-life in people's hepatomicrosome system, and stability is more Good.

Claims (8)

1. the compound shown in a Formulas I or its pharmaceutically acceptable salt:

Wherein R₁、R₂Independently selected from H, D or OH；R₄、R₅、R₆、R₇、R₈、R₉、R₁₀、R₁₁、R₁₂、R₁₃、R₁₄、R₁₅Separately select From H or D, R₃Independently selected from C₁-C₄Alkyl or deuterated C₁-C₄Alkyl, condition is R₁～R₁₅In including at least a D-atom.

2. the compound of claim 1, wherein R₃Independently selected from CH₃、CH₂D、CHD₂Or CD₃。

3. the compound of claim 1, it is selected from following compound:

4. the compound of claim 1, it is selected from following compound:

5. a pharmaceutical composition, its compound comprising claim 1 or its pharmaceutically acceptable salt, and a kind of or many Plant pharmaceutically acceptable carrier, excipient and/or medium.

6. the pharmaceutical composition of compound any one of claim 1-4 or its pharmaceutically acceptable salt, claim 5 exists Purposes in the medicine of the disease that SGLT-2 suppression is benefited from preparation treatment.

7. the purposes of claim 6, the wherein said disease of SGLT-2 suppression of benefiting from is selected from diabetes, diabetic retinal Disease, diabetic neuropathy, diabetic nephropathy, wound healing delay, insulin resistance, hyperglycemia, hyperinsulinism The rising of mass formed by blood stasis, aliphatic acid blood concentration, the rising of glycerine blood concentration, hyperlipidemia, obesity, hypertriglyceridemia, X syndrome, Atherosclerotic or hypertension.

8. the purposes of claim 7, the wherein said disease of SGLT-2 suppression of benefiting from is selected from diabetes.