GSK-525762A

GSK 525762A; 1260907-17-2; I-BET-762; GSK525762A; UNII-5QIO6SRZ2R; 5QIO6SRZ2R;
CAS1260907-17-2
2-[(4S)-6-(4-chlorophenyl)-8-methoxy-1-methyl-4H-[1,2,4]triazolo[4,3-a][1,4]benzodiazepin-4-yl]-N-ethylacetamide

Molecular Formula:	C22H22ClN5O2
Molecular Weight:	423.89538 g/mol

Solubility:	Soluble in DMSO (84 mg/ml at 25 °C), ethanol (42 mg/ml at 25 °C, warmed), DMF (~30 mg/ml), ethanol:PBS (pH 7.2, 1:1) (~0.5 mg/ml), and water (<1 mg/ml at 25 °C).
Storage:	Store at -20° C
Density:	~1.4 g/cm3 (Predicted)
Refractive Index:	n20D 1.67 (Predicted)
Optical Activity:	α20D 85º±5º, c = 0.3 in methanol
IC50:	BRD2: IC50 = 32.5 nM (human); BRD4: IC50 = 36.1 nM (human); BRD3: IC50 = 42.4 nM (human); PBMC: IC50 = 316.23 nM (human); HepG2: EC5050 = 700 nM (human)
pK Values:	pKb: 2.43 (Predicted)

In April 2016, GSK-525762 was reported to be in phase 2 clinical development. GSK-525762 was originally disclosed in WO2011054553, claiming benzodiazepine derivatives as bromodomain inhibitors, useful for treating cancer. See WO2014028547, claiming use of GSK-525762 for treating small cell lung cancer.
GSK 525762A, is a BET Bromodomain Inhibitor, which is now in clinical development. BET bromodomains have emerged as promising drug targets for treatment of cancers, inflammatory diseases, and other medical conditions.
Patent
WO-2016050821
Patent applications WO201 1/054553 and WO201 1/054845 (both in the name of GlaxoSmithKline LLC) disclose the compound 2-[(4S)-6-(4-chlorophenyl)-1-methyl-8-(methyloxy)-4/-/-[1 ,2,4]triazolo[4,3-a][1 ,4]benzodiazepin-4-yl]-/V-ethylacetamide as a BET family bromodomain inhibitor and describes therapeutic uses thereof. The chemical structure of this compound is represented by formula (I):

(I)

Scheme 1




Example 1
Preparation of an acetonitrile solvate of 2-[(4S)-6-(4-chlorophenyl)-1 -methyl-8-(methyloxy)-4H-[1 ,2,4]triazolo[4,3-a][1 ,4]benzodiazepin-4-yl]-yV-ethylacetamide
Amorphous 2-[(4S)-6-(4-chlorophenyl)-1-methyl-8-(methyloxy)-4H-[1 ,2,4]triazolo[4,3-a][1 ,4]benzodiazepin-4-yl]-/V-ethylacetamide (prepared for instance as described in WO201 1/054553, 1 wt) was dissolved in acetonitrile (20 vol) upon heating (up to reflux). The solution was then distilled to 10 vol keeping the temp 50 °C - 60 °C by adjusting the vacuum. Nucleation occurred during the final stage of the distillation. The slurry was then held at 60 °C before being cooled to 20 °C and filtered. The cake was then washed with
acetonitrile (2 vol). The cake was dried under vacuum with a nitrogen bleed at approximately 60 °C to provide the titled product.
¹H-NMR (500 MHz, DMSO-d₆, referenced to TMS = 0.00 ppm, T = 25 C) δ ppm 8.22 (1 H, t, J = 5 Hz), 7.79 (1 H, d, J = 9 Hz), 7.53 (2H, d, J = 9 Hz), 7.49 (2H, d, J = 9 Hz), 7.38 (1 H, dd, J = 3 Hz, 9 Hz), 6.87 (1 H, d, J = 3 Hz), 4.49 (1 H, m), 3.79 (3H, s), 3.25 (1 H, m), 3.20-3.06 (3H, several m), 2.54 (3H, s), 2.08 (3H, s), 1 .07 (3H, t, J = 7 Hz).
Example 2
Preparation of a benzene sulphonic acid salt of 2-[(4S)-6-(4-chlorophenyl)-1 -methyl-8-(methyloxy)-4H-[1 ,2,4]triazolo[4,3-a][1 ,4]benzodiazepin-4-yl]-A/-ethylacetamide in crystalline solid state form
Preparation 1
The acetonitrile solvate of 2-[(4S)-6-(4-chlorophenyl)-1-methyl-8-(methyloxy)-4/-/-[1 ,2,4]triazolo[4,3-a][1 ,4]benzodiazepin-4-yl]-/V-ethylacetamide (for a preparation see Example 1 , 2.58 g) was slurried in acetonitrile (7 mL) and 2-methyltetrahydrofuran (7 mL). Benzenesulfonic acid (1.17 g) was dissolved in acetonitrile (7 mL). The resulting solution was charged to the slurry of 2-[(4S)-6-(4-chlorophenyl)-1-methyl-8-(methyloxy)-4/-/-[1 ,2,4]triazolo[4,3-a][1 ,4]benzodiazepin-4-yl]-/V-ethylacetamide acetonitrile solvate in acetonitrile and 2-methyltetrahydrofuran. An additional rinse of acetonitrile (1.4 mL) and 2-methyltetrahydrufran (0.7 mL) was added to the slurry. The slurry was then warmed to 60 °C to dissolve. 2-methyltetrahydrofuran (50 mL) was then added over 30 minutes. Crystals formed during this addition. The resulting suspension was then cooled to 5 °C at a controlled, linear rate of 0.5 °C/minute. The slurry was aged for 1 hour. The crystalline product was then isolated by filtration and rinsed with a 5 to 1 mixture of 2-methyltetrahydrofuran and acetonitrile (15 mL). The product was then dried in a vacuum oven at 55 °C overnight.
Preparation 2
The acetonitrile solvate of 2-[(4S)-6-(4-chlorophenyl)-1 -methyl-8-(methyloxy)-4/-/-[1 ,2,4]triazolo[4,3-a][1 ,4]benzodiazepin-4-yl]-/V-ethylacetamide (prepared for example in a process such as Example 1 above, 1 wt) was dissolved in 9 vol 2-methyltetrahydrofuran at 65 °C. Once cooled to 20°C the solution was filtered into the crystallization vessel. The dissolution vessel and inline filter were rinsed with 1 vol 2-methyltetrahydrofuran. The solution was then heated to 45 °C.
1 .05 eq of benzene sulphonic acid was dissolved in 1 volume of filtered acetonitrile. 10% of this solution was added to a reactor to which 0.05 wt% of a benzene sulphonic acid
salt of 2-[(4S)-6-(4-chlorophenyl)-1-methyl-8-(methyloxy)-4H-[1 ,2,4]triazolo[4,3-a][1 ,4]benzodiazepin-4-yl]-/V-ethylacetamide micronized seed (prepared for example as in Preparation 1 above) slurry was charged. The remaining benzene sulphonic acid solution was charged at a steady rate over 2 hours, maintaining the reactor at 45 °C.
The slurry was cooled to 0 °C at no greater than 0.2 °C/minute. The slurry was filtered.
The crystallizer was charged with the first wash, 3 vol of filtered 2-methyltetrahydrofuran, which was cooled to <10 °C while stirring in the crystallizer, before being used to wash the cake. The crystallizer was charged with the second wash, 3 vol of filtered 2-methyltetrahydrofuran, which was cooled to <10 °C while stirring in the crystallizer, before being used to wash the cake. The crystallizer was charged with the third wash, 4 vol of filtered 2-MeTHF, which was cooled to <10 °C while stirring in the crystallizer, before being used to wash the cake. The cake was blown-down until the solvent being removed was reduced to a trickle. The title compound was then dried in a vacuum oven at 50 °C until the loss on drying (LOD) indicates <0.2% wt. loss (LOD method: 10 min at 120 °C). The product was then delumped using a comil.
Example 3
Characterisation of a benzene sulphonic acid salt of 2-[(4S)-6-(4-chlorophenyl)-1 -methyl-8-(methyloxy)-4H-[1 ,2,4]triazolo[4,3-a][1 ,4]benzodiazepin-4-yl]-yV-ethyl acetamide in crystalline solid state form
XRPD
The X-ray powder diffraction (XRPD) data were acquired on a PANalytical X'Pert Pro powder diffractometer, model PW3050/60, using an X'Celerator detector. The acquisition conditions were: radiation: Cu Ka, generator tension: 45 kV, generator current: 40 mA, step size: 0.017 °2Θ, time per step: 500 seconds, divergence slit type: fixed, divergence slit size: 0.4354 °, measurement temperature: 20-25 °C, goniometer radius: 240 mm. The sample was prepared by packing sample in a 0.9 mm capillary. Peak positions were obtained using PANalytical X'Pert Highscore Plus software. The margin of error is approximately ± 0.1° 2Θ for each of the peak assignments.
The X-ray powder diffraction (XRPD) pattern is shown in Figure 1 and shows characteristic peaks, expressed in degrees 2Θ, at 5.5, 7.4, 9.1 , 10.0, 10.4, 13.3, 13.6, 14.9, 18.7, 20.4, 20.9, 22.8 and 23.1 ° ( ± 0.1 °).
¹³C Solid State NMR (SSNMR)
A ¹³C SSNMR spectrum was obtained at 273K on a spectrometer operating at a frequency of 100.56 MHz for ¹³C observation using a cross-polarization pulse sequence with a Bruker 4-mm triple resonance magic-angle spinning probe at a rotor frequency of 8 kHz. The margin of error is ± 0.2 ppm for each of the peak assignments.
The ¹³C SSNMR spectrum is shown in Figure 2 and comprises chemical shifts (ppm) at 169.6, 167.5 165.6, 160.1 , 159.4, 157.1 , 155.9, 154.3, 152.4, 146.9, 145.8, 140.0, 137.9, 135.9, 133,4, 132.0, 130.6, 129.9, 128.3, 127.1 , 125.6, 123.5, 120.6, 1 19.1 , 1 14.1 , 1 13.7, 58.0, 53.6, 53.1 , 40.7, 37.0, 34.9, 15.8, 14.7, and 12.0 ( ±0.2 ppm).

PATENT
WO2011054553
http://www.google.com/patents/WO2011054553A1?cl=en
formula (I) which is 2-[(4S)-6-(4- Chlorophenyl)-1-methyl-8-(methyloxy)-4H-[1 ,2,4]triazolo[4,3-a][1 ,4]benzodiazepin-4-yl]-/V- ethylacetamide

(I)
or a salt thereof.
It will be appreciated that the present invention covers compounds of formula (I) as the free base and as salts thereof, for example as a pharmaceutically acceptable salt thereof.
In one embodiment there is provided a compound which is 2-[(4S)-6-(4-Chlorophenyl)-1- methyl-8-(methyloxy)-4H-[1 ,2,4]triazolo[4,3-a][1 ,4]benzodiazepin-4-yl]-/V-ethylacetamide.
Because of their potential use in medicine, salts of the compounds of formula (I) are desirably pharmaceutically acceptable. In another embodiment there is provided a compound which is 2-[(4S)-6-(4-Chlorophenyl)-1-methyl-8-(methyloxy)-4H- [1 ,2,4]triazolo[4,3-a][1 ,4]benzodiazepin-4-yl]-/V-ethylacetamide or a pharmaceutically acceptable salt thereof.
The compound of formula (I) may be prepared according to reaction scheme 1 by reaction of a compound of formula (II) with EtNH₂ in the presence of HATU or HBTU and DIEA at room temperature. Alternatively compounds of formula (I) may be prepared by reacting the compound of formula (II) with oxalyl chloride followed by addition of EtNH₂ in the presence of triethylamine.
Scheme 1

The compound of formula (II) may be prepared according to reaction Scheme 2. Suitable reaction conditions comprise reacting a compound of formula (III) with alkaline hydroxide preferably sodium hydroxide or lithium hydroxide.
Scheme 2

wherein R represents C-|.galkyl such as methyl.
Compounds of formula (III), may be prepared according to reaction scheme 3 by reacting compounds of formula (IV) with AcOH. Scheme 3

Compounds of formula (IV) may be prepared according to reaction scheme 4 by reacting compounds of formula (VI) with hydrazine below 15 °C followed by reaction of the resulting hydrazone (V) with MeCOCI at 0°C. Generally hydrazone (V) is used without further purification and is reacted with MeCOCI at , for example 0 °C.
Scheme 4

(IV) Compounds of formula (VI) in which R is Ci_-6alkyl (such as methyl) may be prepared according to reaction scheme 5 from compounds of formula (VII) by treatment with Lawesson's reagent or P₄Si₀. Suitable reaction conditions comprise reacting compounds of formula (VIII) with P₄Si₀ in 1 ,2-dichloroethane at, for example 70 °C.
Scheme 5

Compounds of formula (VII) may be prepared according to reaction scheme 6, by reacting compounds of formula (IX) with an organic base such as triethylamine followed by reaction of the resulting amine (VIII) with acetic acid. Generally, amine (VIII) is used without further purification and is reacted with AcOH at, for example 60 °C.
Scheme 6

Compounds of formula (IX) may be prepared according to reaction scheme 7, by reacting compounds of formula (XI) with the acylchloride (X) derived from protected aspartic acid. Scheme 7

Compounds of formula (XI) may be prepared according to procedures described in Synthesis 1980, 677-688. Acyl chlorides of formula (X) may be prepared according to procedures described in J. Org. Chem., 1990, 55, 3068-3074 and J. Chem. Soc. Perkin Trans. 1 , 2001 , 1673-1695.
Alternatively the compound of formula (I) may be prepared according to reaction scheme 8.

wherein R represents C-|_4alkyl such as methyl.
The compound of formula (IIIA) may be prepared according to reaction scheme 9 by reacting compounds of formula (IVA) with EtNH₂ in the presence of HATU and DIEA at, for example room temperature.
Scheme 9

The compound of formula (IVA) may be prepared according to reaction scheme 10. Suitable reaction conditions comprise reacting compounds of formula (VI) with alkaline hydroxide such as sodium hydroxide. Scheme 10

Example 1 : 2-[(4S)-6-(4-Chlorophenyl)-1 -methyl-8-(methyloxy)-4H-[1 ,2,4]triazolo[4,3-

To a solution of [(4S)-6-(4-Chlorophenyl)-1-methyl-8-(methyloxy)-4H-[1 _!2_!4]triazolo[4,3- a][1 ,4]benzodiazepin-4-yl]acetic acid (for a preparation see Intermediate 1 )(16.0 g, 40 mmol) in THF at RT was added DIEA (14 mL, 80 mmol) followed by HATU (30.4 g, 80 mmol). The reaction mixture was stirred for 3h at this temperature and ethylamine (40 mL, 2M in THF, 80 mmol) was added. The mixture was stirred for 48h before being concentrated under reduced pressure. The crude material was suspended in water and extracted with DCM. The organic layer was dried over Na₂S0₄, filtered and concentrated in vacuo. The crude solid was purified by chromatography on Si0₂ (DCM/MeOH 95/5) and the resulting solid recrystallised in MeCN. The solid was then dissolved in DCM and precipited with /^'-Pr₂0 to give the title compound (8 g, 47% yield) as a white solid.
R_f = 0.48 (DCM/MeOH : 90/10). Mp >140 °C (becomes gummy). ¹H NMR (300 MHz, CDCI₃) 7.53-7.47 (m, 2H), 7.39 (d, J = 8.9 Hz, 1 H), 7.37-7.31 (m, 2H), 7.20 (dd, J = 2.9 and 8.9 Hz, 1 H), 6.86 (d, J = 2.9 Hz, 1 H), 6.40 (m, 1 H), 4.62 (m, 1 H), 3.80 (s, 3H), 3.51 (dd, J = 7.3 and 14.1 Hz, 1 H), 3.46-3.21 (m, 3H), 2.62 (s, 3H), 1.19 (t, J = 7.3 Hz, 3H). LC/MS : m/z 424 [M(³⁵CI)+H]⁺, Rt 2.33 min.
Intermediate 1 : [(4S)-6-(4-Chlorophenyl)-1 -methyl-8-(methyloxy)-4H-
[1 ,2,4]triazolo[4,3-a][1 ,4]benzodiazepin-4-yl]acetic acid

To a solution of methyl [(4S)-6-(4-chlorophenyl)-1 -methyl-8-(methyloxy)-4H- [1 ,2,4]triazolo[4,3-a][1 ,4]benzodiazepin-4-yl]acetate (for a preparation see Intermediate 2)(28 g, 68 mmol) in THF (450 mL) at RT was added 1 N NaOH (136 mL, 136 mmol). The reaction mixture was stirred at this temperature for 5h before being cooled down and quenched with 1 N HCI (136 mL). THF was removed under reduced pressure and the aqueous layer was extracted with DCM. The combined organic layers were dried over Na₂S0₄, filtered and concentrated under reduced pressure. The crude solid was recrystallised in CH₃CN to give the title compound (23.9 g, 89% yield) as a pale yellow powder. ¹H NMR (300 MHz, CDCI₃) δ 7.55-7.48 (m, 2H), 7.41 (d, J = 8.9 Hz, 1 H), 7.38- 7.31 (m, 2H), 7.22 (dd, J = 2.9 and 8.9 Hz, 1 H), 6.90 (d, J = 2.9 Hz, 1 H), 4.59 (dd, J = 6.9 and 6.9 Hz, 1 H), 3.81 (s, 3H), 3.70 (dd, J = 6.9 and 25.7 Hz, 1 H), 3.61 (dd, J = 6.9 and 25.7 Hz, 1 H), 2.63 (s, 3H). LC/MS: m/z 397 [M(³⁵CI)+H]⁺, Rt 2.1 1 min.
Intermediate 2: Methyl [(4S)-6-(4-chlorophenyl)-1 -methyl-8-(methyloxy)-4H- [1 ,2,4]triazolo[4,3-a][1 ,4]benz

To crude methyl [(3S)-2-[(1 Z)-2-acetylhydrazino]-5-(4-chlorophenyl)-7-(methyloxy)-3H- 1 ,4-benzodiazepin-3-yl]acetate (for a preparation see Intermediate 3) (34 g, 79 mmol) was suspended in THF (200 mL) and AcOH (200 mL) was added at RT. The reaction mixture was stirred at this temperature overnight before being concentrated to dryness. The residue was suspended in saturated NaHC0₃ and extracted with DCM. The organic layer was dried over Na₂S0₄, filtered and concentrated in vacuo. The crude solid was purified by chromatography on Si0₂ (DCM/MeOH : 90/10) to give the title compound (28 g, 86% yield) as a yellow powder.
1H NMR (300 MHz, CDCI₃) δ 7.54-7.47 (m, 2H), 7.40 (d, J = 8.8 Hz, 1 H), 7.37-7.31 (m, 2H), 7.22 (dd, J = 2.8 and 8.8 Hz, 1 H), 6.89 (d, J = 2.8 Hz, 1 H), 4.61 (dd, J = 6.4 and 7.8 Hz, 1 H), 3.82 (s, 3H), 3.78 (s, 3H), 3.66 (dd, J = 7.8 and 16.9 Hz, 1 H), 3.60 (dd, J = 6.4 and 16.9 Hz, 1 H), 2.62 (s, 3H). LC/MS m/z 41 1 [M(³⁵CI)+H]⁺, Rt 2.88 min. Intermediate 3: Methyl [(3S)-2-[2-acetylhydrazino]-5-(4-chlorophenyl)-7-(methyloxy)- 3H-1 ,4-benzodiazepin-3-yl]acetate

To a suspension of methyl [(3S)-5-(4-chlorophenyl)-7-(methyloxy)-2-thioxo-2,3-dihydro- 1 H-1 ,4-benzodiazepin-3-yl]acetate (for a preparation see Intermediate 4)(30.2 g, 77.7 mmol) in THF (800 mL) at 0°C was added hydrazine monohydrate (1 1 .3 ml_, 233 mmol) dropwise. The reaction mixture was stirred for 4h between 0°C and 15°C before being cooled at 0°C. Et₃N (32.4 mL, 230 mmol) was then added slowly and AcCI (16.3 mL, 230 mmol) was added dropwise. The mixture was allowed to warm to RT and stir for 1 h then quenched with water and concentrated under reduced pressure. The resulting aqueous layer was then extracted with DCM and the organic layer was dried over Na₂S0₄, filtered and concentrated in vacuo to give the crude title compound (34 g, 100% yield) which was used without further purification. LC/MS: m/z 429 [M(³⁵CI)+H]⁺, Rt 2.83 min. Intermediate 4: Methyl [(3S)-5-(4-chlorophenyl)-7-(methyloxy)-2-thioxo-2,3-dihydro- 1H-1 ,4-benzodiazepin-3-yl]acetate

A suspension of P₄S_i0 (85.8 g, 190 mmol) and Na₂C0₃ (20.5 g, 190 mmol) in 1 ,2-DCE (1.5 L) at RT was stirred for 1 h before methyl [(3S)-5-(4-chlorophenyl)-7-(methyloxy)-2- oxo-2,3-dihydro-1 H-1 ,4-benzodiazepin-3-yl]acetate (for a preparation see Intermediate 5) (40 g, 107 mmol) was added. The resulting mixture was stirred at 65°C for 4 h before being cooled and filtered. The solid was washed with DCM and the filtrate washed with sat. NaHC0₃. The organic layer was dried over Na₂S0₄, filtered and concentrated under reduced pressure. The title compound was precipitated from a DCM//^'-Pr₂0 mixture and filtered. The filtrate was then concentrated and purified by flash chromatography (DCM/MeOH : 98/2) to afford another batch of product. The title compound was obtained combining the two fractions (30.2 g, 73%) as a yellow powder. LC/MS: m/z 389
[M(³⁵CI)+H]⁺, Rt 3.29 min.
Intermediate 5: Methyl [(3S)-5-(4-chlorophenyl)-7-(methyloxy)-2-oxo-2,3-dihydro-1H- 1 ,4-benzodiazepin-3-yl]acetat

To a solution of the crude methyl /V¹-[2-[(4-chlorophenyl)carbonyl]-4-(methyloxy)phenyl]- /V²-{[(9H-fluoren-9-ylmethyl)oxy]carbonyl}-L-a-asparaginate (for a preparation see Intermediate 6) (assumed 0.2 mol) in DCM (500 mL) was added Et₃N (500 mL, 3.65 mol) and the resulting mixture was refluxed for 24h before being concentrated. The resulting crude amine was dissolved in 1 ,2-DCE (1.5 L) and AcOH (104 mL, 1.8 mol) was added carefully. The reaction mixture was then stirred at 60°C for 2h before being concentrated in vacuo and dissolved in DCM. The organic layer was washed with 1 N HCI and the aqueous layer was extracted with DCM (x3). The combined organic layers were washed twice with water, and brine, dried over Na₂S0₄, filtered and concentrated under reduced pressure. The crude solid was recrystallised in MeCN leading to the title compound (51 g) as a pale yellow solid. The filtrate could be concentrated and recrystallised in MeCN to give another 10 g of Intermediate 9 (total: 61 g, 69% yield based on recovered
Intermediate 12). R_f = 0.34 (DCM/MeOH : 95/5). LC/MS m/z 373 [M(³⁵CI)+H]⁺, Rt 2.76 min.
Intermediate 6: Methyl W^2-[(4 :hlorophenyl)carbonyl]-4-(methyloxy)phenyl] V²- {[(9H-fluoren-9-ylmethyl)oxy]carbonyl}-L-a-asparaginate

A mixture of Methyl /V-{[(9H-fluoren-9-ylmethyl)oxy]carbonyl}-L-a-aspartyl chloride (prepared from J. Org. C em. 1990, 55, 3068-3074 and J. C em. Soc. Perkin Trans. 1 2001 , 1673-1695) (221 g, 0.57 mol) and [2-amino-5-(methyloxy)phenyl](4- chlorophenyl)methanone (for a preparation see Intermediate 7) (133 g, 0.5 mol) in CHCI₃ (410 mL) was stirred at 60°C for 1.5h before being cooled and concentrated under reduced pressure and used without further purification. LC/MS: m/z 613 [M(³⁵CI)+H]⁺, Rt = 3.89 min. Intermediate 7: [2-amino-5-(methyloxy)phenyl](4-chlorophenyl)methanone

To a solution of 2-methyl-6-(methyloxy)-4H-3,1-benzoxazin-4-one (for a preparation see Intermediate 8)(40.0 g, 0.21 mol) in a toluene (560 ml_)/ether (200 mL) mixture at 0°C was added dropwise a solution of 4-chlorophenylmagnesium bromide (170 mL, 1 M in Et₂0, 0.17 mol). The reaction mixture was allowed to warm to RT and stirred for 1 h before being quenched with 1 N HCI. The aqueous layer was extracted with EtOAc (3 x) and the combined organics were washed with brine, dried over Na₂S0₄, filtered and concentrated under reduced pressure. The crude compound was then dissolved in EtOH (400 mL) and 6N HCI (160 mL) was added. The reaction mixture was refluxed for 2 h before being concentrated under reduced pressure. The resulting solid was filtered and washed twice with ether before being suspended in EtOAc and neutralised with 1 N NaOH. The aqueous layer was extracted with EtOAc (3 x) and the combined organics were washed with brine, dried over Na₂S0₄, filtered and concentrated under reduced pressure. The title compound was obtained as a yellow solid (39 g, 88 % yield) which was used without further purification. Intermediate 8 : 2-methyl-6-(methyloxy)-4H-3,1 -benzoxazin-4-one

A solution of 5-methoxyanthranilic acid (7.8 g, 46.5 mmol) was refluxed in acetic anhydride (60 mL) for 2h15 before being cooled and concentrated under reduced pressure. The crude residue was then concentrated twice in the presence of toluene before being filtered and washed with ether to yield to the title compound (6.8 g, 77% yield) as a beige solid; LC/MS: m/z 192 [M+H]⁺, Rt 1.69 min.
Preparation of reference compound for use in biological assays
Experimental details of LC-MS methods A and B as referred to herein are as follows:
LC/MS (Method A) was conducted on a Supelcosil LCABZ+PLUS column (3μηΊ, 3.3cm x 4.6mm ID) eluting with 0.1 % HCO2H and 0.01 M ammonium acetate in water (solvent A), and 95% acetonitrile and 0.05% HCO2H in water (solvent B), using the following elution gradient 0-0.7 minutes 0%B, 0.7-4.2 minutes 0→100%B, 4.2-5.3 minutes 100%B, 5.3-5.5 minutes 100→0%B at a flow rate of 3 mL/minute. The mass spectra (MS) were recorded on a Fisons VG Platform mass spectrometer using electrospray positive ionisation [(ES+ve to give [M+H]⁺ and [M+NH4]⁺ molecular ions] or electrospray negative ionisation
[(ES-ve to give [M-H]- molecular ion] modes. Analytical data from this apparatus are given with the following format : [M+H]⁺ or [M-H]-.
LC/MS (Method B) was conducted on an Sunfire C18 column (30mm x 4.6mm i.d. 3.5μηι packing diameter) at 30 degrees centigrade, eluting with 0.1 % v/v solution of Trifluoroacetic Acid in Water (Solvent A) and 0.1 % v/v solution of Trifluoroacetic Acid in Acetonitrile (Solvent B) using the following elution gradient 0-0.1 min 3%B, 0.1- 4.2min 3 - 100% B, 4.2-4.8min 100% B, 4.8-4.9min 100-3%B, 4.9 - 5.0min 3% B at a flow rate of 3ml/min. The UV detection was an averaged signal from wavelength of 210nm to 350nm and mass spectra were recorded on a mass spectrometer using positive electrospray ionization. Ionisation data was rounded to the nearest integer. LC/HRMS: Analytical HPLC was conducted on a Uptisphere-hsc column (3μηι 33 x 3 mm id) eluting with 0.01 M ammonium acetate in water (solvent A) and 100% acetonitrile (solvent B), using the following elution gradient 0-0.5 minutes 5% B, 0.5-3.75 minutes 5→100% B, 3.75-4.5 100% B, 4.5-5 100→5% B, 5-5.5 5% B at a flow rate of 1 .3 mL/minute. The mass spectra (MS) were recorded on a micromass LCT mass spectrometer using electrospray positive ionisation [ES+ve to give MH⁺ molecular ions] or electrospray negative ionisation [ES-ve to give (M-H)- molecular ions] modes.
TLC (thin layer chromatography) refers to the use of TLC plates sold by Merck coated with silica gel 60 F254.
Silica chromatography techniques include either automated (Flashmaster or Biotage SP4) techniques or manual chromatography on pre-packed cartridges (SPE) or manually- packed flash columns.
Reference compound A : 2-meth -6-(methyloxy)-4H-3,1 -benzoxazin-4-one

A solution of 5-methoxyanthranilic acid (Lancaster) (41.8 g, 0.25 mol) was refluxed in acetic anhydride (230 mL) for 3.5 h before being concentrated under reduced pressure. The crude compound was then concentrated twice in the presence of toluene before being filtered and washed twice with ether to yield to the title compound (33.7 g, 71 % yield) as a brown solid; LC/MS (Method A): m/z 192 [M+H]⁺, Rt 1.69 min.
Reference compound B: [2-amino- -(methyloxy)phenyl](4-chlorophenyl)methanone

To a solution of 2-methyl-6-(methyloxy)-4H-3,1-benzoxazin-4-one (for a preparation see Reference compound A) (40.0 g, 0.21 mol) in a toluene/ether (2/1 ) mixture (760 mL) at 0°C was added dropwise a solution of 4-chlorophenylmagnesium bromide (170 mL, 1 M in Et₂0, 0.17 mol). The reaction mixture was allowed to warm to room temperature and stirred for 1 h before being quenched with 1 N HCI (200 mL). The aqueous layer was extracted with EtOAc (3 x 150 mL) and the combined organics were washed with brine (100 mL), dried over Na₂S0₄, filtered and concentrated under reduced pressure. The crude compound was then dissolved in EtOH (400 mL) and 6N HCI (160 mL) was added. The reaction mixture was refluxed for 2 h before being concentrated to one-third in volume. The resulting solid was filtered and washed twice with ether before being suspended in EtOAc and neutralised with 1 N NaOH. The aqueous layer was extracted with EtOAc (3 x 150 mL) and the combined organics were washed with brine (150 mL), dried over Na₂S0₄, filtered and concentrated under reduced pressure. The title compound was obtained as a yellow solid (39 g, 88 % yield); LC/MS (Method A): m/z 262 [M+H]+, Rt 2.57 min.
Reference Compound C: Methyl /^-^-[(^chlorophenyljcarbonyl]^- (methyloxy)phenyl]-yV²-{[(9H-fluoren-9-ylmethyl)oxy]carbonyl}-L-a-asparaginate

Methyl /V-{[(9H-fluoren-9-ylmethyl)oxy]carbonyl}-L-a-aspartyl chloride {Int. J. Peptide Protein Res. 1992, 40, 13-18) (93 g, 0.24 mol) was dissolved in CHCI₃ (270 mL) and [2- amino-5-(methyloxy)phenyl](4-chlorophenyl)methanone (for a preparation see Reference compound B) (53 g, 0.2 mol) was added. The resulting mixture was stirred at 60°C for 1 h before being cooled and concentrated at 60% in volume. Ether was added at 0°C and the resulting precipitate was filtered and discarded. The filtrate was concentrated under reduced pressure and used without further purification.
Reference compound D: Methyl [(3S)-5-(4-chlorophenyl)-7-(methyloxy)-2-oxo-2,3- dihydro-1H-1 ,4-benzodiazepin-3-yl]acetate

To a solution of Methyl N1-[2-[(4-chlorophenyl)carbonyl]-4-(methyloxy)phenyl]-N2-{[(9H- fluoren-9-ylmethyl)oxy]carbonyl}-L-a-asparaginate (for a preparation see Reference compound C) (assumed 0.2 mol) in DCM (500 mL) was added Et₃N (500 mL, 3.65 mol) and the resulting mixture was refluxed for 24h before being concentrated. The resulting crude amine was dissolved in 1 ,2-DCE (1.5 L) and AcOH (104 mL, 1.8 mol) was added carefully. The reaction mixture was then stirred at 60°C for 2h before being concentrated in vacuo and dissolved in DCM. The organic layer was washed with 1 N HCI and the aqueous layer was extracted with DCM (x3). The combined organic layers were washed twice with water, and brine, dried over Na₂S0₄, filtered and concentrated under reduced pressure. The crude solid was recrystallised in MeCN leading to the title compound (51 g) as a pale yellow solid. The filtrate could be concentrated and recrystallised in MeCN to give to another 10 g of the desired product R_f = 0.34 (DCM/MeOH : 95/5).
HRMS (M+H)⁺ calculated for C₁₉H₁₈ ³⁵CIN₂0₄ 373.0955; found 373.0957.
Reference compound E: Methyl [(3S)-5-(4-chlorophenyl)-7-(methyloxy)-2-thioxo-2,3- dihydro-1 H-1 ,4-benzodiazepi -3-yl]acetate

A suspension of P₄S_i0 (36.1 g, 81.1 mmol) and Na₂C0₃ (8.6 g, 81.1 mmol) in 1 ,2-DCE (700 mL) at room temperature was stirred for 2 h before Methyl [(3S)-5-(4-chlorophenyl)- 7-(methyloxy)-2-oxo-2,3-dihydro-1 H-1 ,4-benzodiazepin-3-yl]acetate (for a preparation see Reference compound D) (16.8 g, 45.1 mmol) was added. The resulting mixture was stirred at 70°C for 2 h before being cooled and filtered. The solid was washed twice with DCM and the filtrate washed with sat. NaHC0₃ and brine. The organic layer was dried over Na₂S0₄, filtered and concentrated under reduced pressure. The crude product was purified by flash-chromatography on silica gel (DCM/MeOH : 99/1 ) to afford the title compound (17.2 g, 98% yield) as a yellowish solid. LC/MS (Method A): m/z 389 [M(³⁵CI)+H]⁺, Rt 2.64 min
HRMS (M+H)⁺ calculated for C₁₉H₁₈ ³⁵CIN₂0₃S 389.0727; found 389.0714. Reference compound F: Methyl [(3S)-2-[2-acetylhydrazino]-5-(4-chlorophenyl)-7- (methyloxy)-3H-1 ,4-benzodiazepin-3- l]acetate

To a suspension of Methyl [(3S)-5-(4-chlorophenyl)-7-(methyloxy)-2-thioxo-2,3-dihydro- 1 H-1 ,4-benzodiazepin-3-yl]acetate (for a preparation see Reference compound E (9.0 g, 23.2 mmol) in THF (300 mL) at 0°C was added hydrazine monohydrate (3.4 mL, 69.6 mmol) dropwise. The reaction mixture was stirred for 5h between 5°C and 15°C before being cooled at 0°C. Et₃N (9.7 mL, 69.6 mmol) was then added slowly and acetyl chloride (7.95 mL, 69.6 mmol) was added dropwise. The mixture was then allowed to warm to room temperature for 16h before being concentrated under reduced pressure. The crude product was dissolved in DCM and washed with water. The organic layer was dried over Na₂S0₄, filtered and concentrated in vacuo to give the crude title compound (9.7 g, 98% yield) which was used without further purification. R_f = 0.49 (DCM/MeOH : 90/10).
Reference compound G: Methyl [(4S)-6-(4-chlorophenyl)-1 -methyl-8-(methyloxy)-4H- [1 ,2,4]triazolo[4,3-a][1 ,4]benz

The crude Methyl [(3S)-2-[(1 Z)-2-acetylhydrazino]-5-(4-chlorophenyl)-7-(methyloxy)-3H- 1 ,4-benzodiazepin-3-yl]acetate (for a preparation see Reference compound F) (assumed 9.7 g) was suspended in THF (100 ml) and AcOH (60 mL) was added at room temperature. The reaction mixture was stirred at this temperature for 2 days before being concentrated under reduced pressure. The crude solid was triturated in /^'-Pr₂0 and filtered to give the title compound (8.7 g, 91 % over 3 steps) as an off-white solid.
HRMS (M+H)⁺ calculated for C21 H2₀CIN4O₃ 41 1.1229; found 41 1.1245.
Reference compound H: [(4S)-6-(4-Chlorophenyl)-1 -methyl-8-(methyloxy)-4H- [1 ,2,4]triazolo[4,3-a][1 ,4]benzodiazepin-4-yl]acetic acid

To a solution of Methyl [(4S)-6-(4-chlorophenyl)-1 -methyl-8-(methyloxy)-4H- [1 ,2,4]triazolo[4,3-a][1 ,4]benzodiazepin-4-yl]acetate (for a preparation see Reference compound G)(7.4 g, 18.1 mmol) in THF (130 mL) at room temperature was added 1 N NaOH (36.2 mL, 36.2 mmol). The reaction mixture was stirred at this temperature for 5h before being quenched with 1 N HCI (36.2 mL) and concentrated in vacuo. Water is then added and the aqueous layer was extracted with DCM (x3) and the combined organic layers were dried over Na₂S0₄, filtered and concentrated under reduced pressure to give the title compound (7 g, 98% yield) as a pale yellow solid.
PATENT
WO2014028547


http://www.nature.com/nature/journal/v468/n7327/fig_tab/nature09589_F1.html



http://www.google.com/patents/WO2014028547A1?cl=en
Zhao, Y., et al.: J. Med. Chem., 56, 7498 (2013); Mirguet, O., et al.: J. Med. Chem., 56, 7501 (2013);

Patent ID	Date	Patent Title
US2015119435	2015-04-30	COMPOUNDS USEFUL FOR PROMOTING PROTEIN DEGRADATION AND METHODS USING SAME
US8975417	2015-03-10	Pyrazolopyrrolidine derivatives and their use in the treatment of disease
US2014356322	2014-12-04	Compounds & Methods for the Enhanced Degradation of Targeted Proteins & Other Polypeptides by an E3 Ubiquitin Ligase
US2014243322	2014-08-28	BIVALENT BROMODOMAIN LIGANDS, AND METHODS OF USING SAME
US2014243321	2014-08-28	BIOORTHOGONAL MONOMERS CAPABLE OF DIMERIZING AND TARGETING BROMODOMAINS, AND METHODS OF USING SAME
US2014243286	2014-08-28	BROMODOMAIN LIGANDS CAPABLE OF DIMERIZING IN AN AQUEOUS SOLUTION, AND METHODS OF USING SAME
US2013079335	2013-03-28	Benzotriazolodiazepine Compounds Inhibitors Of Bromodomains
US2012252781	2012-10-04	Benzodiazepine Bromodomain Inhibitor
US2012220573	2012-08-30	Benzodiazepine Bromodomain Inhibitor
US2012208800	2012-08-16	Bromodomain Inhibitors For Treating Autoimmune And Inflammatory Diseases

Patent ID	Date	Patent Title
US2015366877	2015-12-24	METHODS OF TREATMENT OF HUMAN CYTOMEGALOVIRUS INFECTION AND DISEASES WITH BROMODOMAIN INHIBITORS
US2015299210	2015-10-22	Benzodiazepine Bromodomain Inhibitor
US2015291562	2015-10-15	IMIDE-BASED MODULATORS OF PROTEOLYSIS AND ASSOCIATED METHODS OF USE
US2015246923	2015-09-03	9H-PYRIMIDO[4,5-B]INDOLES AND RELATED ANALOGS AS BET BROMODOMAIN INHIBITORS
US2015238507	2015-08-27	BENZODIAZEPINES FOR TREATING SMALL CELL LUNG CANCER
US2015210706	2015-07-30	Benzodiazepine Bromodomain Inhibitor
US2015174138	2015-06-25	DIAGNOSTIC AND TREATMENT METHODS IN SUBJECTS HAVING OR AT RISK OF DEVELOPING RESISTANCE TO CANCER THERAPY
US2015133436	2015-05-14	Bromodomain Inhibitors For Treating Autoimmune And Inflammatory Diseases
US2015133434	2015-05-14	Compositions and Methods for Reactivating Latent Immunodeficiency Virus
US2015119435	2015-04-30	COMPOUNDS USEFUL FOR PROMOTING PROTEIN DEGRADATION AND METHODS USING SAME

/////GSK-525762A, GSK-525762, GSK 525762A, GSK 525762, 1260907-17-2, phase 2,
CCNC(=O)CC1C2=NN=C(N2C3=C(C=C(C=C3)OC)C(=N1)C4=CC=C(C=C4)Cl)C

TAK-058 (ENV-8058)

TAK-058 , ENV-8058
5-HT 3 receptor antagonist
Envoy Therapeutics, Inc.
1-(1-methyl-1H-pyrazol-4-yl)-N-((1R,5S,7S)-9-methyl-3-oxa-9-azabicyclo[3.3.1]nonan-7-yl)-1H-indole-3-carboxamide
l-(l-methyl-lH-pyrazol-4-yl)-N-((lR,5 .7S)-9-methyl-3-oxa-9-azabicyclo[3.3.11nonan-7-yl)-lH-indole-3-carboxamide
1-(1-methyl-1H- pyrazol-4-yl)-N- ((1R,5S,7S)- 9-methyl-3- oxa-9-azabicyclo [3.3.1]nonan-7- yl)-1H-indole-3- carboxamide, 2,2,2- trifluoroacetic acid salt
N-(9-methyl-3-oxa-9-azabicyclo[3.3.1]nonan-7-yl)-1-(1-methylpyrazol-4-yl)indole-3-carboxamide

Molecular Formula:	C21H25N5O2
Molecular Weight:	379.4555 g/mol

https://clinicaltrials.gov/ct2/show/NCT02153099
Phase I Schizophrenia

Company	Takeda Pharmaceutical Co. Ltd.
Description	Serotonin (5-HT3) receptor antagonist
Molecular Target	Serotonin (5-HT3) receptor
Mechanism of Action	Serotonin (5-HT3) receptor antagonist
Therapeutic Modality	Small molecule

Latest Stage of Development	Phase I
Standard Indication	Schizophrenia
Indication Details	Treat schizophrenia

• 01 Dec 2015 Phase-I clinical trials in Schizophrenia (Combination therapy) in USA (PO)
• 01 Dec 2015 Takeda completes a phase I trial in Healthy volunteers in USA (NCT02389881)
• 28 Nov 2015 Takeda plans a phase I trial in Schizophrenia (Combination therapy) in USA (NCT02614586)

1 -( 1 -methyl- 1 H-pyrazol-4-yl)-N-((lR,5S,7S)-9-methyl-3-oxa-9-azabicyclo[3.3.1]nonan-7-yl)-lH-indole-3-carboxamide, free base, which is an antagonist of the 5-HT3 receptor. 1 -(1 -Methyl- 1 H-pyrazol-4-yl)-N-((lR,5S,7S)-9-methyl-3-oxa-9-azabicyclo[3.3.1]nonan-7-yl)-lH-indole-3-carboxamide, 2,2,2-trifluoroacetic acid salt, is disclosed in PCT Publication No. WO
2014/014951, published January 23, 2014.

1-(1-methyl-1H-pyrazol-4-yl)-N-((1R,5S,7S)-9-methyl-3-oxa-9-azabicyclo[3.3.1]nonan-7-yl)-1H-indole-3-carboxamide a 5-HT3 receptor antagonist, useful for treating anxiety, depression, eating disorder, schizophrenia, cognitive dysfunction, Parkinson's disease, Huntington's Chorea, presenile dementia, Alzheimer's disease and atherosclerosis.
This compound was originally claimed in WO2014014951,  Takeda, following its acquisition of Envoy Therapeutics, is developing TAK-058 (ENV-8058), a 5-HT3 receptor antagonist, as an oral solution for treating schizophrenia, especially cognitive impairment associated with schizophrenia.
In July 2015, the drug was listed as being in phase I development. TAK-058 may have emerged from a schizophrenia therapy program which used Envoy's bacTRAP translational profiling technology to identify a protein target in the brain.
PATENT
WO2014014951
Example 5
Synthesis of l-(l-methyl-lH-pyrazol-4-yl)-N-((lR,5 .7S)-9-methyl-3-oxa-9-azabicyclo[3.3.11nonan-7-yl)-lH-indole-3-carboxamide. 2.2.2-trifluoroacetic acid salt

Step 1 : methyl 1-(1 -methyl- lH-pyrazol-4-yl)-lH-indole-3-carboxylate. TFA
To a sealed tube was added copper(I) iodide (65.2 mg, 0.342 mmol), methyl 1H-indole-3-carboxylate (200 mg, 1.142 mmol) and potassium phosphate (509 mg, 2.397 mmol), then the reaction vessel was evacuated and purged with nitrogen (3x). Next, 4-bromo-l-methyl-lH-pyrazole (184 mg, 1.142 mmol) and (lR,2R)- ,N2-dimethylcyclohexane-l,2-diamine (109 μΐ, 0.685 mmol) were added, followed by toluene (1 142 μΐ). The reaction tube was evacuated and purged with nitrogen, then sealed and heated at 1 10 °C for 24 h. HPLC purification provided the title compound as a colorless oil.
Step 2: 1-(1 -methyl- lH-pyrazol-4-yl)-lH-indole-3-carboxylic acid hydrochloride
To a solution of methyl 1-(1 -methyl- lH-pyrazol-4-yl)-lH-indole-3-carboxylate, TFA
(3.5 mg, 9.48 μιηοΐ) in MeOH (95 μΐ) was added a solution of aq. KOH (33.2 μΐ, 0.066 mmol, 2 M). The reaction mixture was stirred at RT overnight, then acidified with IN HC1.
The solvent was evaporated under reduced pressure and the residue was dried under vacuum overnight. The title compound was used without further purification.
Step 3 : l-(l-methyl-lH-pyrazol-4-yl)-N-((lR,5 .7S)-9-methyl-3-oxa-9-azabicyclor3.3.11nonan-7-yl)-lH-indole-3-carboxamide, 2,2,2-trifluoroacetic acid salt
To a mixture of 1-(1 -methyl- lH-pyrazol-4-yl)-lH-indole-3-carboxylic acid hydrochloride (2.6 mg, 9.36 μιηοΐ) in DMF (187 μΐ) was added HATU (4.27 mg, 0.01 1 mmol) and DIPEA (8.18 μΐ, 0.047 mmol). After the reaction mixture was stirred at RT for 15 min, (lR,5S,7S)-9-methyl-3-oxa-9-azabicyclo[3.3.1]nonan-7-amine, TFA (3.04 mg, 0.01 1 mmol) was added and stirring was continued for 2 h. HPLC purification afforded the title compound as a white solid. MS (ESI, pos. ion) m/z: 380.30 (M+l).

PATENT
WO-2016053947
EXAMPLE 1 : l-(l-methyl-lH-pyrazol-4-yl)-N-((lR,5S,7S)-9-methyl-3-oxa-9-azabicyclo[3.3.1 ]nonan-7-yl)- lH-indole-3-carboxamide
l-(l-Methyl-lH-pyrazol-4-yl)-lH-indole-3-carboxylic acid (128.7 g, 0.53 mol,) and anhydrous THF (645 mL) was heated to about 43°C. Oxalyl chloride (137.7 g, 92 mL, 1.08 mol) was added dropwise between 40 and 50°C. Gas evolution ceased in approximately 30 minutes. The resulting suspension was stirred for 2 hours at 50°C, allowed to cool to room temperature, and then stirred overnight. The suspension was diluted with heptane (1.5 L), stirred for 10 minutes, and allowed to settle. The supernatant was removed. The addition of heptane (1.5 L), followed by stirring, settling, and decanting was repeated two more times.
The resulting suspension was diluted with anhydrous THF (645 mL) and the ratio between THF and heptane was determined by NMR to be 3:2. The reaction mixture was cooled to 5°C and to the mixture was added DIPEA base (138 g, 1.07 mol) at such a rate that the temperature did not exceed 20°C. Next (li?,55^*,7S)-9-methyl-3-oxa-9-azabicyclo[3.3.1]nonan-7-amine (101.4 g, 0.63 mol) in 500 mL of anhydrous THF was added. The reaction mixture was warmed to ambient temperature and stirred at 20 to 23°C overnight to give a suspension.
The suspension was filtered and the cake was dissolved in IN HC1 (2.6 L). The aqueous layer was washed with EtOAc (3 x 2.6 L). The aqueous layer was cooled to 5°C and was basified to pH 12 with aqueous potassium hydroxide (230 g) solution in water (500 mL). The mixture was stirred at 5 to 10°C overnight to give a solid. The product was filtered, washed with water (2 x 1.2 L), followed by MTBE (2 x 1.2 L), and then dried to give 128 g (64%) of the (crude) title compound.
Patent
https://www.google.co.in/patents/US20140024644
1-(1-methyl-1H- pyrazol-4-yl)-N- ((1R,5S,7S)- 9-methyl-3- oxa-9-azabicyclo [3.3.1]nonan-7- yl)-1H-indole-3- carboxamide, 2,2,2- trifluoroacetic acid salt
Synthetic Procedures Reference 1 Synthesis of (1R,5S,7S)-tert-butyl 7-hydroxy-3-oxa-9-azabicyclo[3.3.1]nonane-9-carboxylate

•  
  
•  
  Sodium borohydride (259 mg, 6.84 mmol) was added portion-wise to a solution of (1R,5S)-tert-butyl 7-oxo-3-oxa-9-azabicyclo[3.3.1]nonane-9-carboxylate (550 mg, 2.279 mmol) in MeOH (4559 μl) at 0° C. After 5 min, the reaction mixture was allowed to warm to RT then stirred for 30 min. The mixture was concentrated under reduced pressure, dissolved in EtOAc and washed with brine. The combined organic layers were dried over anhydrous Na₂SO₄, filtered and concentrated under reduced pressure to afford the title compound as a white solid, which was used without further purification.

Example 4 Synthesis of N-((1R,5S,7S)-9-methyl-3-oxa-9-azabicyclo[3.3.1]nonan-7-yl)-1-(1H-pyrazol-4-yl)-1H-indole-3-carboxamide, 2,2,2-trifluoroacetic acid salt

•  
  
•  
  A mixture of 1-((1-benzyl-1H-pyrazol-4-yl)-N-((1R,5S,7S)-9-methyl-3-oxa-9-azabicyclo[3.3.1]nonan-7-yl)-1H-indole-3-carboxamide 2,2,2-trifluoroacetate (85 mg, 0.149 mmol) and 10% Pd—C (120 mg) in MeOH (1.0 ml) was stirred at RT under H₂ for 2 days. Filtration and concentration afforded the title compound as a white solid. MS (ESI, pos. ion) m/z: 366.20 (M+1).

Example 5 Synthesis of 1-(1-methyl-1H-pyrazol-4-yl)-N-((1R,5S,7S)-9-methyl-3-oxa-9-azabicyclo[3.3.1]nonan-7-yl)-1H-indole-3-carboxamide, 2,2,2-trifluoroacetic acid salt

•  
  

Step 1: methyl 1-(1-methyl-1H-pyrazol-4-yl)-1H-indole-3-carboxylate, TFA

•  
  To a sealed tube was added copper(I) iodide (65.2 mg, 0.342 mmol), methyl 1H-indole-3-carboxylate (200 mg, 1.142 mmol) and potassium phosphate (509 mg, 2.397 mmol), then the reaction vessel was evacuated and purged with nitrogen (3×). Next, 4-bromo-1-methyl-1H-pyrazole (184 mg, 1.142 mmol) and (1R,2R)—N1,N2-dimethylcyclohexane-1,2-diamine (109 μl, 0.685 mmol) were added, followed by toluene (1142 μl). The reaction tube was evacuated and purged with nitrogen, then sealed and heated at 110° C. for 24 h. HPLC purification provided the title compound as a colorless oil.

Step 2: 1-(1-methyl-1H-pyrazol-4-yl)-1H-indole-3-carboxylic acid hydrochloride

•  
  To a solution of methyl 1-(1-methyl-1H-pyrazol-4-yl)-1H-indole-3-carboxylate, TFA (3.5 mg, 9.48 μmol) in MeOH (95 μl) was added a solution of aq. KOH (33.2 μl, 0.066 mmol, 2 M). The reaction mixture was stirred at RT overnight, then acidified with 1N HCl. The solvent was evaporated under reduced pressure and the residue was dried under vacuum overnight. The title compound was used without further purification.

Step 3: 1-(1-methyl-1H-pyrazol-4-yl)-N-((1R,5S,7S)-9-methyl-3-oxa-9-azabicyclo[3.3.1]nonan-7-yl)-1H-indole-3-carboxamide, 2,2,2-trifluoroacetic acid salt

•  
  To a mixture of 1-(1-methyl-1H-pyrazol-4-yl)-1H-indole-3-carboxylic acid hydrochloride (2.6 mg, 9.36 μmol) in DMF (187 μl) was added HATU (4.27 mg, 0.011 mmol) and DIPEA (8.18 μl, 0.047 mmol). After the reaction mixture was stirred at RT for 15 min, (1R,5S,7S)-9-methyl-3-oxa-9-azabicyclo[3.3.1]nonan-7-amine, TFA (3.04 mg, 0.011 mmol) was added and stirring was continued for 2 h. HPLC purification afforded the title compound as a white solid. MS (ESI, pos. ion) m/z: 380.30 (M+1).

15

TFA

379.456 MW

380.30  MS +1

Patent ID	Date	Patent Title
US2015182533	2015-07-02	5-HT3 RECEPTOR ANTAGONISTS
US2014024644	2014-01-23	5-HT3 RECEPTOR ANTAGONISTS

 

/////////TAK-058 , ENV-8058, phase I, takeda, 5-HT 3 receptor antagonist, Envoy Therapeutics, Inc., Phase I,  Schizophrenia
C12CC(CC(N1C)COC2)NC(c4c3ccccc3n(c4)c5cnn(c5)C)=O
CN1C=C(C=N1)N2C=C(C3=CC=CC=C32)C(=O)NC4CC5COCC(C4)N5C

Henagliflozin

Henagliflozin, SHR-3824 ,
CAS 1623804-44-3
C22-H24-Cl-F-O7
454.8756
PHASE 2 for the treatment of type 2 diabetes
HengRui (Originator)

Jiangsu Hengrui Medicine Co Ltd

UNII-21P2M98388; 21P2M98388; Henagliflozin; SHR3824; SHR-3824;
In April 2016, Jiangsu Hengrui Medicine is developing henagliflozin (phase 2 clinical trial), a sodium-glucose cotransporter-2 (SGLT-2) inhibitor, for treating type 2 diabetes. 
SGLT1 and SGLT2 inhibitors, useful for treating eg diabetes.
Henagliflozin proline is in phase II clinical trials by Jiangsu Hengrui (江苏恒瑞) for the treatment of type 2 diabetes.
1,6-dehydrated-1-C{4-chloro-3-[(3-fluoro-4-ethoxyphenyl)methyl]phenyl}-5-C-(hydroxymethyl)-β-L-idopyranose L-proline
(1 ^ 2345-5- [4-chloro-3 - [(4-ethoxy-3-fluoro-phenyl) - methyl] phenyl] -1- (hydroxymethyl) 6,8 - alcohol dioxide
(1R,2S,3S,4R,5R)-5-[4-chloro-3-[(4-ethoxy-3-fluorophenyl)methyl]phenyl]-1-(hydroxymethyl)-6,8-dioxabicyclo[3.2.1]octane-2,3,4-triol

Shanghai Hengrui Pharmaceutical Co., Ltd., 上海恒瑞医药有限公司, Jiangsu Hengrui Medicine Co., Ltd., 江苏恒瑞医药股份有限公司, Less «

• 01 May 2015 Jiangsu HengRui Medicine Co. initiates enrolment in a phase I drug interaction trial in volunteers in China (NCT02500485)
• 12 Feb 2015 Jiangsu HengRui Medicine plans a phase I trial for Type-2 diabetes mellitus in China (NCT02366377)
• 01 Feb 2015 Jiangsu HengRui Medicine initiates enrolment in a phase I trial for Type-2 diabetes mellitus in China (NCT02366351)

Henagliflozin is a novel sodium-glucose transporter 2 inhibitor and presents a complementary therapy to metformin for patients with T2DM due to its insulin-independent mechanism of action. This study evaluated the potential pharmacokinetic drug-drug interaction between henagliflozin and metformin in healthy Chinese male subjects. 2. In open-label, single-center, single-arm, two-period, three-treatment self-control study, 12 subjects received 25 mg henagliflozin, 1000 mg metformin or the combination. Lack of PK interaction was defined as the ratio of geometric means and 90% confidence interval (CI) for combination: monotherapy being within the range of 0.80-1.25. 3. Co-administration of henagliflozin with metformin had no effect on henagliflozin area under the plasma concentration-time curve (AUC_0-24) (GRM: 1.08; CI: 1.05, 1.10) and peak plasma concentration (C_max) (GRM: 0.99; CI: 0.92, 1.07). Reciprocally, co-administration of metformin with henagliflozin had no clinically significant on metformin AUC_0-24 (GRM: 1.09, CI: 1.02, 1.16) although there was an 11% increase in metformin C_max (GRM 1.12; CI 1.02, 1.23). All monotherapies and combination therapy were well tolerated. 4. Henagliflozin can be co-administered with metformin without dose adjustment of either drug.

PATENT
WO-2016050134

With the improvement of socio-economic development and living standards, worldwide rapid growth of diabetes, diabetes is usually divided into two kinds of diabetes type Ⅰ and type Ⅱ diabetes, more than 90% of type Ⅱ diabetes. Species has been listed diabetes drugs a lot, but so far, no drugs which can single-handedly blood glucose levels in patients with type Ⅱ diabetes in the long-term target range. In recent years, in-depth study of the pathogenesis of diabetes, for the treatment of type Ⅱ diabetes provide more and more ways, and sodium - glucose cotransporter 2 (sodium-glucose transporter 2, SGLT-2) inhibitors found for treatment of diabetes provides another new idea. SGLT-2 inhibitors in the treatment mechanism of inhibition of SGLT-2 activity by selective to lower blood sugar. Select the SGLT-2 as a target, partly because of its absolute weight of glucose absorption, and partly because it is only expressed in the kidney. The current study also found that the mechanism of SGLT-2 does not depend on the degree of abnormal function of β cells or insulin resistance, its effect is not as severe failure or insulin resistance and β-cell function decline.Therefore, it is reasonable that the SGLT-2 inhibitors for the treatment of type Ⅱ diabetes currently has good prospects.

WO2012019496 discloses SGLT-2 inhibitor of the following formula, and its chemical name is 1,6-anhydro -1-C- {4- chloro-3 - [(3-fluoro-4-ethoxyphenyl) methyl] phenyl} -5-C- (hydroxymethyl) -β-L- idose pyranose.

However, direct 1,6-anhydro -1-C- {4- chloro-3 - [(3-fluoro-4-ethoxyphenyl) methyl] phenyl} -5-C- (hydroxymethyl) - β-L- idose pyranose as a pharmaceutically active ingredient is not realistic, because a lower melting point (83 ℃), having a hygroscopicity, poor development of the form, therefore, to develop it into a stable form of the compound having the transformation very important.

Example 1

Take (1.0g, 2.2mmol) 1,6- dehydration -1-C- {4- chloro-3 - [(3-fluoro-4-ethoxyphenyl) methyl] phenyl} -5-C- ( hydroxymethyl) -β-L- Aidoo pyranose (prepared by the method disclosed in WO2012019496), in 7.20g ethanol addition was completed, stirring to dissolve. Was added at room temperature L- proline (0.2786g, 2.42mmol, 1.1eq), the addition was completed, the reaction was warmed at reflux for 10min, the reaction solution was clear, hot filtered and the filtrate was stirred to room temperature, there is a lot of white solid precipitated , allowed to stand overnight, filtered, and dried, to give the formula (I), compound as a white solid 1.14 g, yield 88%. X- ray diffraction spectrum of the crystalline sample is shown in Figure 1. The crystallization at about 5.41 (16.33) 7.69 (11.49), 10.22 (8.65) 12.04 (7.35), 12.46 (7.10), 14.42 (6.14), 17.30 (5.12), 18.79 (4.72), 19.38 (4.58), 20.24 (4.38), 22.73 (3.91), 24.58 (3.62), 27.55 (3.24), 28.82 (3.10) and 31.03 (2.88) at the characteristic peaks. DSC spectrum shown in Figure 2, has a melting endothermic peak 111.20 ℃, this is defined as a Form A polymorph.

PATENT
WO2012019496
https://www.google.com/patents/WO2012019496A1?cl=en
Example 4
(1 ^ 2345-5- [4-chloro-3 - [(4-ethoxy-3-fluoro-phenyl) - methyl] phenyl] -1- (hydroxymethyl) 6,8 - alcohol dioxide

first step
1-ethoxy-2-fluoro - benzene
A mixture of 2-fluoro-phenol 4a (6.7 g, 60 mmol) was dissolved in 66 mL of acetone, was added iodoethane (6.3 mL,
78 mmol) and potassium carbonate (12.4 g, 90 mmol), at reflux in an oil bath for 5 hours. The reaction solution was concentrated under reduced pressure, was added 100 mL of ethyl acetate and 60 mL of water, separated, the aqueous phase was extracted with ethyl acetate (30 mLx2), the organic phases combined, dried over anhydrous magnesium sulfate, filtered, and the filtrate was concentrated under reduced pressure, to give the title product 1-ethoxy-2-fluoro - benzene 4b (6.9 g, red oil). yield: 82.1%.
MS m / z (ESI): 280.2 [2M + 1]
The second step
(5-bromo-2-chloro - phenyl) - (4-ethoxy-3-fluoro-phenyl) - methanone A mixture of 5-bromo-2-chloro - benzoyl chloride 2a (12.4 g, 48.8 mmol) was dissolved a 100 mL of dichloromethane was added 1-ethoxy-2-fluoro - benzene 4b (6.84 g, 48.8 mmol), cooled to 0 ° C, was added portionwise aluminum (5.86 g, 44 mmol) chloride, 16 h. Was added dropwise under ice-cooling to the reaction mixture 20 mL of 2 M HCl solution, separated, the aqueous phase was extracted with 30 mL of dichloromethane, and the combined organic phase was dried over anhydrous magnesium sulfate, filtered, and the filtrate was concentrated under reduced pressure to give the title The product (5-bromo-2-chloro - phenyl) - (4-ethoxy-3-fluoro-phenyl) - methanone 4c (12.7 g, yellow solid), yield: 72.6%.
MS m / z (ESI): 358.9 [M + l] Step
₍₅ - bromo-2-chloro - phenyl) - (4-ethoxy-3-fluoro-phenyl) - methanol (5-Bromo-2-chloro - phenyl) - (4-ethoxy -3 - fluoro - phenyl) -methanone 4c (12.7 g, 35.5 mmol) was dissolved in methanol and a 100 mL of tetrahydrofuran (ν: ν = 1: 1) mixed solvent, under an ice bath was added portionwise sodium borohydride (2.68 g, 70 mmol), and reacted at room temperature for 30 minutes. Add 15 mL of acetone, the reaction solution was concentrated under reduced pressure, 150 mL of ethyl acetate was added to dissolve the residue, washed with saturated sodium chloride solution (50 mLx2). The combined organic phase was dried over anhydrous magnesium sulfate, filtered, and concentrated under reduced pressure The filtrate, to give the title product (5-bromo-2-chloro - phenyl) - (4-ethoxy-3-fluoro-phenyl) - methanol 4d (12.7 g, orange oil), was used directly without isolation next reaction.
the fourth step
4 - [(5-bromo-2-chloro-phenyl) - methyl] Small-ethoxy-2-fluoro - benzene (5-bromo-2-chloro - phenyl) - (4-ethoxy -3 - fluoro - phenyl) methanol 4d (12.7 g, 35.3 mmol) was dissolved in a 100 mL of dichloromethane was added triethylsilane (16.9 mL, 106 mmol), was added dropwise boron trifluoride etherate (8.95 mL, 70.6 mmol ), for 3 hours. Was added 50 mL of saturated sodium bicarbonate solution, separated, the aqueous phase was extracted with ethyl acetate (100 mLx2), the organic phases combined, dried over anhydrous magnesium sulfate, filtered, and the filtrate was concentrated under reduced pressure, purified by silica gel column chromatography to elute B surfactant system resulting residue was purified to give the title product 4 - [(5-bromo-2-chloro - phenyl) methyl] -1-ethoxy-2-fluoro - benzene 4e (10 g, as a pale yellow oil ) yield: 82.4%.
_{1H NMR (400 MHz, CDC1 3} ): δ 7.33-7.27 (m, 3H), 6.95-6.90 (m, 3H), 4.14 (q, 2H), 4.01 (s, 2H), 1.49 (t, 3H)
the fifth step
(2 3R, 4S, 5 ^ 6R) -2- [4- chloro-3 - [(4-ethoxy-3-fluoro-phenyl) - methyl] phenyl] -6- (hydroxymethyl) - 2-methoxy - tetrahydro-pyran-3,4,5-triol
4 - [(5-bromo-2-chloro - phenyl) methyl] -1-ethoxy-2-fluoro - benzene 4e (7.36 g, 21.4 mmol) was dissolved in 30 mL of tetrahydrofuran, cooled to -78 ° C, was added dropwise a solution of n-butyllithium in hexane (10.27 mL, 25.7 mmol), at -78 ° C to react 1 hour, a solution of 20 mL (3R, 4S, 5R, 6R) -3,4,5 - tris (trimethylsilyloxy) -6- (trimethylsilyloxy) tetrahydropyran-2-one 2f (llg, 23.6 mmol) in tetrahydrofuran at -78 ° C under reaction 2 h, 2.8 mL of methanesulfonic acid and 71 mL of methanol, the reaction at room temperature for 16 hours. Was added 100 mL of saturated sodium carbonate solution, the reaction solution was concentrated under reduced pressure, to the residue was added 50 mL of saturated sodium chloride solution, extracted with ethyl acetate (100 mLx3), organic phases were combined, dried over anhydrous magnesium sulfate, filtered, The filtrate was concentrated under reduced pressure, purified by silica gel column chromatography with eluent systems resulting A residue was purified to give the title product (2 3R, 4S, 5 6R) -2- [4- chloro-3 - [(4-ethoxyphenyl 3-fluoro-phenyl) - methyl] phenyl] -6- (hydroxymethyl) -2-methoxy - tetrahydro-pyran-3,4,5-triol 4f (5.7 g, white solid ) yield: 58.3%.
_{1H NMR (400 MHz, CD 3} OD): δ 7.56 (s, 1H), 7.48 (dd, 1H), 7.37 (dd, 1H), 6.95-6.87 (m, 3H), 4.08-4.07 (m, 4H) , 3.91 (m, 1H), 3.93-3.73 (m, 2H), 3.56-3.53 (m, 1H), 3.45-3.43 (m, 1H), 3.30 (s, 2H), 3.08 (s, 3H), 1.35 (t, 3H)
The sixth step
(2 3R, 4S, 5 6R) -6- [(tert-butyl (dimethyl) silyl) oxymethyl] -2- [4-chloro-3 - [(4-ethoxy-3-fluoro - phenyl) methyl] phenyl] -2-methoxy - tetrahydro-pyran-3,4,5-triol the (2 3R, 4S, 5 6R) -2- [4- chloro-3- [(4-ethoxy-3-fluoro-phenyl) - methyl] phenyl] -6- (hydroxymethyl) -2-methoxy - 4f tetrahydropyran-3,4,5-triol (5.7 g, 12.5 mmol) was dissolved in 50 mL of pyridine, followed by adding tert-butyldimethylsilyl chloride (2.26 g, 15 mmol) and 4-dimethylaminopyridine (305 mg, 2.5 mmol), for 16 hours. The reaction solution was concentrated under reduced pressure, was added 200 mL of ethyl acetate, washed with a saturated copper sulfate solution (50 mLx3). The combined organic phase was dried over anhydrous magnesium sulfate, filtered, and the filtrate was concentrated under reduced pressure to give the title product (2 3R, 4S, 5 6R) -6- [(tert-butyl (dimethyl) silyl) oxymethyl] -2- [4-chloro-3 - [(4-ethoxy-3-fluoro-phenyl) - methyl] phenyl] -2-methoxy - tetrahydro-pyran-3,4,5-triol 4g (7.14 g, colorless oil), without isolation directly used for the next reaction.
Seventh Step
[[(2R, 3R, 4S, 5R, 6 ^ -3,4,5- tris-benzyloxy-6- [4-chloro-3 - [(4-ethoxy-3-fluoro-phenyl) - methyl yl] phenyl] -6-methoxy - tetrahydropyran-2-yl] methoxy] - tert-butyl - dimethyl-silane (2 3R, 4S, 5 6R) -6- [(tert butyl (dimethyl) silyl) oxymethyl] -2- [4-chloro-3 - [(4-ethoxy-3-fluoro-phenyl) - methyl] phenyl] -2-methoxy yl - tetrahydro-pyran-3,4,5-triol 4g (7.14 g, 12.5 mmol) was dissolved in 100 mL N, N- dimethylformamide was added 60% sodium hydride under ice-cooling (2.5 g , 62.5 mmol), and reacted at room temperature for 40 minutes completed the opening force, was added benzyl bromide (7.5 mL, 62.5 mmol), reaction of 16 hours. 20 mL of methanol, the reaction solution was concentrated under reduced pressure, was added 200 mL of ethyl acetate and 50 mL of water to dissolve the residue, separated, the aqueous phase was extracted with ethyl acetate (50 mL), the organic phase was washed with water (50 mL), washed with saturated sodium chloride solution (50 mL), the combined organic phase was dried over anhydrous magnesium sulfate , filtered, and the filtrate was concentrated under reduced pressure to give the title product [[(2R, 3R, 4S, 5R, 6 ^ -3,4,5- tris-benzyloxy-6- [4-chloro-3 - [(4- ethoxy-3-fluoro-phenyl) - methyl] phenyl] -6-methoxy - tetrahydropyran-2-yl] methoxy] - tert-butyl - dimethylsilane 4h (10.5 g , yellow oil) yield: 99.8%.
Step Eight
[(2R, 3R, 4S, 5R, 6 -3,4,5- tris-benzyloxy-6- [4-chloro-3 - [(4-ethoxy-3-fluoro-phenyl) - methyl] phenyl] -6-methoxy - tetrahydropyran-2-yl] methanol
The [[(2R, 3R, 4S, 5R, 6 -3,4,5- tris-benzyloxy-6- [4-chloro-3 - [(4-ethoxy-3-fluoro-phenyl) - methyl yl] phenyl] -6-methoxy - tetrahydropyran-2-yl] methoxy] - tert-butyl - dimethylsilane 4h (10.52 g, 12.5 mmol) was dissolved in 50 mL of methanol dropwise add acetyl chloride CO.13 mL, 1.9 mmol), for 1 hour. The reaction solution was concentrated under reduced pressure, purified by silica gel column chromatography with eluent systems B resultant residue was purified to give the title product [(2R, 3R, 4S, 5R, 6 -3,4,5- tris-benzyloxy--6 - [4-chloro-3 - [(4-ethoxy-3-fluoro-phenyl) - methyl] phenyl] -6-methoxy - tetrahydropyran-2-yl] methanol 4i (7.6 g , yellow oil yield: 83.6%.
Step Nine
(2 ^ 3456 3,4,5-tris-benzyloxy-6- [4-chloro-3 - [(4-ethoxy-3-fluoro-phenyl) - methyl] phenyl] - 6-methoxy - tetrahydropyran-2-carbaldehyde
Oxalyl chloride (1.17 mL, 13.6 mmol) was dissolved in 20 mL of dichloromethane, cooled to -78 ° C, were added dropwise 20 mL of dimethyl sulfoxide (1.56 mL, 21.9 mmol) in methylene chloride and 50 mL [(2R, 3R, 4S, 5R, 6 -3,4,5- tris-benzyloxy-6- [4-chloro-3 - [(4-ethoxy-3-fluoro-phenyl) - methyl] phenyl] -6-methoxy - tetrahydropyran-2-yl] methanol 4i (7.6 g, 10.45 mmol) in methylene chloride, and reacted at -78 ° C for 30 min, triethylamine (7.25 mL, 52.3 mmol), 2 hours at room temperature was added 50 mL 1 M HCl solution, separated, the organic phase was washed with saturated sodium chloride solution (50 mL x 2), the aqueous phase was extracted with dichloromethane (50 mL), the combined organic phase was dried over anhydrous magnesium sulfate, filtered, and the filtrate was concentrated under reduced pressure to give the title product (2 ^ 3456 3,4,5-tris-benzyloxy-6- [4-chloro-3 - [(4 - ethoxy-3-fluoro-phenyl) - methyl] phenyl] -6-methoxy - tetrahydropyran-2-carbaldehyde 4j (7.58 g, colorless oil), was used directly without isolation next reaction.
The tenth step
(2S, 3 4S, 5R, 6 -3,4,5- tris-benzyloxy-6- [4-chloro-3 - [(4-ethoxy-3-fluoro-phenyl) - methyl] phenyl ] -2- (hydroxymethyl) -6-methoxy - tetrahydropyran-2-carbaldehyde
The (23456 3,4,5-tris-benzyloxy-6- [4-chloro-3 - [(4-ethoxy-3-fluoro-phenyl) - methyl] phenyl] - 6-methoxy - tetrahydropyran-2-carbaldehyde 4j (7.6 g, 10.45 mmol) was dissolved in 80 mL 1,4- dioxane, followed by adding 15.8 mL 37% aqueous formaldehyde and sodium hydroxide solution (31.35 mL, 31.35 mmol), reacted at 70 ° C for 16 h. Add 50 mL of saturated sodium chloride solution, extracted with ethyl acetate (50 mLx4), the organic phase was washed with saturated sodium bicarbonate solution (50 mL), washed with saturated sodium chloride solution (50 mL), the combined organic phase was dried over anhydrous magnesium sulfate, filtered, and the filtrate was concentrated under reduced pressure to give the title product (23,456 benzyloxy-3,4,5-tris - 6- [4-chloro-3 - [(4-ethoxy-3-fluoro-phenyl) - methyl] phenyl] -2- (hydroxymethyl) -6-methoxy - tetrahydropyran - 2- formaldehyde _{4k (7.9g,} as a colorless oil), without isolation directly used for the next reaction.
Step Eleven
[(3 4S, 5R, 6 -3,4,5- tris-benzyloxy-6- [4-chloro-3 - [(4-ethoxy-3-fluoro-phenyl) - methyl] phenyl] 2- (hydroxymethyl) -6-methoxy - tetrahydropyran-2-yl] methanol
The (23456 3,4,5-tris-benzyloxy-6- [4-chloro-3 - [(4-ethoxy-3-fluoro-phenyl) - methyl] phenyl] - 2- (hydroxymethyl) -6-methoxy - tetrahydropyran-2-carbaldehyde 4k (7.9 g, 10.45 mmol) was dissolved in 50 mL of tetrahydrofuran and methanol (v: v = 2: 3) mixed solvent , was added sodium borohydride (794 mg, 20.9 mmol), for 30 minutes. Add a small amount of acetone, the reaction solution was concentrated under reduced pressure, purified by silica gel column chromatography with eluent systems resulting A residue was purified to give the title product, 5R, 6 -3,4,5-tris-benzyloxy-6- [4-chloro-3 - [(4-ethoxy-3-fluoro-phenyl) - methyl] phenyl] -2- (hydroxymethyl ) -6-methoxy - tetrahydropyran-2-yl] methanol 4m (l.ll g, colorless oil). yield: 14.1%.
Step Twelve
[(12345 ^ -2,3,4-tris-benzyloxy-5- [4-chloro-3 - [(4-ethoxy-3-fluoro-phenyl) - methyl] phenyl] 6,8-dioxa-bicyclo [3.2.1] octane-1-yl] methanol
The [(3S, 4S, 5R, 6 -3,4,5- tris-benzyloxy-6- [4-chloro-3 - [(4-ethoxy-3-fluoro-phenyl) - methyl] benzene yl] -2- (hydroxymethyl) -6-methoxy - tetrahydropyran-2-yl] methanol 4m (l.ll g, 1.46 mmol) was dissolved in 20 mL of dichloromethane, cooled to -10 ° C, was added trifluoroacetic acid (0.23 mL, 3 mmol), and reacted at room temperature for 2 hours. 20 mL of saturated sodium bicarbonate solution, separated, the aqueous phase was extracted with dichloromethane (20 mL> <2), and the combined organic phase was dried over anhydrous magnesium sulfate, filtered, and the filtrate was concentrated under reduced pressure, purified by silica gel column chromatography with eluent systems B resultant residue was purified to give the title product [(1 2 3 4R, 5 -2,3,4- tris-benzyloxy-5- [4-chloro-3 - [(4-ethoxy-3-fluoro-phenyl) - methyl] phenyl] 6,8-dioxa-bicyclo [3.2.1] octane-1-yl] methanol 4nC830 mg, colorless oil). yield: 78.3%.
MS m / z (ESI): 742.3 [M + 18]
Thirteenth Step
(12345-5- [4-chloro-3 - [(4-ethoxy-3-fluoro-phenyl) - methyl] phenyl] -1- (hydroxymethyl) -6,8 dioxa-bicyclo [3.2.1] octane-2,3,4-triol
The [(1 2 3 4R, 5S) -2,3,4- tris-benzyloxy-5- [4-chloro-3 - [(4-ethoxy-3-fluoro-phenyl) - methyl] benzene yl] -6,8-dioxa-bicyclo [3.2.1] octane-1-yl] methanol 4n (830 mg, 1.14 mmol) was dissolved in 20 mL of tetrahydrofuran and methanol (v: v = l: l) the a mixed solvent of o-dichlorobenzene was added (1.3 mL, 1 1.4 mmol) and Pd / C (500 mg, 10%), purged with hydrogen three times, the reaction for 3 hours. The reaction solution was filtered, rinsed with a small amount of ethyl acetate, the filtrate was concentrated under reduced pressure, purified by silica gel column chromatography with eluent systems resulting A residue was purified to give the title product (1S, 2 3S, 4R, 5 -5- [ 4-chloro-3 - [(4-ethoxy-3-fluoro-phenyl) - methyl] phenyl] -1- (hydroxymethyl) -6,8-dioxa-bicyclo [3.2.1] octane-2,3,4-triol 4 (420 mg, white solid), yield: 81.0% MS m / z (ESI):. 472.2 [m + 18]
_{1H NMR (400 MHz, CD 3} OD): δ 7.47 (s, 1H), 7.42-7.35 (m, 2H), 6.95-6.87 (m, 3H), 4.16-4.14 (m, 1H), 4.06-4.02 ( m, 4H), 3.85-3.70 (m, 2H), 3.67-3.54 (m, 4H), 1.37 (t, 3H)

////////Henagliflozin, SHR-3824 , PHASE 2,  type 2 diabete,  UNII-21P2M98388,  21P2M98388,  SHR 3824,  SHR3824,

CCOc1ccc(cc1F)Cc2cc(ccc2Cl)[C@]34[C@@H]([C@H]([C@@H]([C@](O3)(CO4)CO)O)O)O