CN110092803A - Tenofovir Chinese mugwort draws the preparation and its application of phenol amine fumarate process impurity - Google Patents

Tenofovir Chinese mugwort draws the preparation and its application of phenol amine fumarate process impurity Download PDF

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CN110092803A
CN110092803A CN201810099676.9A CN201810099676A CN110092803A CN 110092803 A CN110092803 A CN 110092803A CN 201810099676 A CN201810099676 A CN 201810099676A CN 110092803 A CN110092803 A CN 110092803A
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base
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oxygen
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贾慧娟
陈岩
任晓慧
武凯洋
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Beijing Creatron Institute Of Pharmaceutical Research Co Ltd
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Abstract

It ends the present invention relates to a kind of tenofovir and draws the process impurity and preparation method thereof of phenol amine fumarate 9- [(R) -2- [[(S)-[[(S) -1- (isopropoxy carbonyl) ethyl] amino] phenoxy group phosphinyl] methoxyl group] propyl] adenine hemifumarate, wherein the impurity is selected from compound A, (S)-isopropyl 2- (((R)-((((R) -1- (6- amino -9H-9- purine radicals) -2- propyl) oxygen) methyl) (methoxyl group) phosphinyl) amino) propionic ester fumarate etc..

Description

Tenofovir Chinese mugwort draws the preparation and its application of phenol amine fumarate process impurity
Technical field
The invention belongs to pharmaceutical synthesis fields, are related to process impurity and its preparation during production of raw medicine, in particular to Tenofovir Chinese mugwort draws phenol amine fumarate process impurity and preparation method thereof.
Background technique
Hepatitis B has become global public health problem.There are about 2,000,000,000 people to infect hepatitis B in the whole world Malicious (HBV), has 500,000~700,000 people to die of hepatic failure caused by HBV infection, cirrhosis and hepatocellular carcinoma every year.China's chronic The number about 1.3 hundred million of Hepatitis virus infection, there are a large amount of patients because of acute or chronic hepatitis B in studies have shown that China every year Cause dead.
Treating the crucial of the disease at present is still antiviral therapy, and antiviral drugs is divided into two major classes, i.e. ucleosides and dry Plain class is disturbed, wherein nucleoside medicine is more extensive in clinical treatment, and drug improves patient by the inhibition to virus replication Liver function, help to improve life in patients.This kind of drug includes: 1998 by Food and Drug Adminstration of the US (FDA) The cytosine nucleoside reverse transcriptase inhibitor Lamivudine of the GlaxoSmithKline PLC of approval；The granted adenine core of in September, 2002 Glycoside reverse transcriptase inhibitor Aldoforwe ester；Obtain within 2005 the guanosint glycoside reverse transcription of FDA approval Shi Guibao company Enzyme inhibitor Entecavir；Granted tenofovir disoproxil fumarate in 2008.
It is the tenofovir disoproxil fumarate of a upgrade version that tenofovir Chinese mugwort, which draws phenol amine fumarate, and drug effect is replaced with fumaric acid Nuo Fuwei ester is identical, but dosage and toxicity are far smaller than tenofovir disoproxil fumarate, and bioavilability is high.
Tenofovir Chinese mugwort draws the chemical name of phenol amine fumarate are as follows: 9- [(R) -2- [[(S)-[[(S) -1- (isopropoxy Carbonyl) ethyl] amino] phenoxy group phosphinyl] methoxyl group] propyl] adenine hemifumarate, structural formula are as follows:
Tenofovir Chinese mugwort draws the synthetic route of phenol amine fumarate to report in patent CN103842366, the plan of the route Slightly: with (R) -9- [2- (phosphate methoxy) propyl] adenine (SM1) and triphenyl phosphite (SM3) for starting material, Under the action of triethylamine, esterification is carried out in acetonitrile, obtains intermediate ((((R) -1- (6- amino -9H- purine -9- base) Propane -2- base) oxygroup) methyl) phosphoric acid monophenyl (TAF-1).The intermediate, with thionyl chloride heating stirring, obtains in toluene Phosphinylidyne chlorization product (TAF-2).TAF-2 using with l-Alanine isopropyl ester (SM2) be condensed, obtain 9- [(R) -2- [[(R, S)-[[(S) -1- (isopropoxy carbonyl) ethyl] amino] phenoxy group phosphinyl] methoxyl group] propyl] adenine fumarate, it should Fumarate obtains 9- [(R) -2- [[(R, S)-[[(S) -1- (isopropoxy carbonyl) second at salt with fumaric acid again after free Base] amino] phenoxy group phosphinyl] methoxyl group] propyl] adenine hemifumarate (TAF).
The route is the synthetic strategy with industrial production value.Currently, there is no to the technique generated in this route The report of impurity.
Summary of the invention
Impurity research is that the important content of drug research and development always through drug research and development directly affects the safety of drug Property, validity.Phenol amine fumarate and corresponding formulation products quality research is drawn to provide impurity control in order to end to tenofovir Product improve tenofovir Chinese mugwort and draw phenol amine Related product quality standard, the safe medication of phenol amine fumarate is drawn for tenofovir Chinese mugwort Important guidance is provided, present invention research synthesizes and confirmed that tenofovir Chinese mugwort draws the work in phenol amine fumarate production line Skill impurity.
The tenofovir Chinese mugwort of patent CN103842366 report is drawn in the synthetic method of phenol amine fumarate, and a system can be generated The process impurity of column, chemical name and structural formula see the table below:
The way of production of the above impurity is as follows:
Compound A, compound B: in step 1, last handling process needs to use methanol, under alkaline condition, with system In intermediate TAF-1 ester exchange reaction occurs, generate impurity intermediate, and further across step 2 and step 3, transmitting Into finished product, compound A, B are generated.The differentiation foundation of compound A, B: two compounds are by TAF-3 in methanol solution Transesterification preparation, judges from mechanism, it should the overturning of phosphorus chirality occurs, therefore production quantity is more for compound A, it is less to be Compound B.
Compound C, compound D: in step 1, treatment process also needs to use ethyl alcohol, and ester occurs with intermediate TAF-1 and hands over Reaction is changed, impurity intermediate is generated, and then pass through step 2 and step 3, is transmitted in finished product, generate compound C, D.Change Close the differentiation foundation of object C, D: by TAF-3 prepared by the transesterification in ethanol solution for two compounds, judges from mechanism, it should The overturning of phosphorus chirality occurs, therefore production quantity is more for compound C, it is less for compound D.
Compound E, compound F: in intermediate TAF-1, methanol can occur as residual solvent with TAF-2 effect Esterification generates impurity intermediate, and then passes through step 3, is transmitted in finished product, generates compound E, F；Compound E, F Differentiation foundation: analogy tenofovir Chinese mugwort draws the phosphine chiral selectivity of phenol amine, and in compound E, F, phosphine chirality is based on S configuration Product is wanted, therefore, production quantity is more for compound E, less for compound F.
Compound G, compound H: in intermediate TAF-1, ethyl alcohol can occur as residual solvent with TAF-2 effect Esterification generates impurity intermediate, and then passes through step 3, is transmitted in finished product, generates compound G, H.Compound G, H Differentiation foundation: analogy tenofovir Chinese mugwort draws the phosphine chiral selectivity of phenol amine, and in compound G, H, phosphine chirality is based on S configuration Product is wanted, therefore, production quantity is more for compound G, less for compound H.
Compound I contains impurity asparagus fern door propylhomoserin isopropyl ester in starting material 2 (l-Alanine isopropyl ester), is transmitted to end In product, compound I is generated.
As a result, on one side, the present invention relates to compound A, its chemical name is: (S)-isopropyl 2- (((R)- ((((R) -1- (6- amino -9H-9- purine radicals) -2- propyl) oxygen) methyl) (methoxyl group) phosphinyl) amino) propionic ester fumaric acid Salt, structural formula are as follows:
On the other hand, the present invention relates to compound B, its chemical name is: (S)-isopropyl 2- (((S)-((((R) -1- (6- amino -9H-9- purine radicals) -2- propyl) oxygen) methyl) (methoxyl group) phosphinyl) amino) propionic ester fumarate, structure Formula is as follows:
On the other hand, the present invention relates to compound C, its chemical name is: (S)-isopropyl 2- (((R)-((((R) -1- (6- amino -9H-9- purine radicals) -2- propyl) oxygen) methyl) (ethyoxyl) phosphinyl) amino) propionic ester fumarate, structure Formula is as follows:
On the other hand, the present invention relates to compound D, its chemical name is: (S)-isopropyl 2- (((S)-((((R) -1- (6- amino -9H-9- purine radicals) -2- propyl) oxygen) methyl) (ethyoxyl) phosphinyl) amino) propionic ester fumarate, structure Formula is as follows:
On the other hand, the present invention relates to compound E, its chemical name is: (S)-aminomethyl phenyl ((((R) -1- (6- amino - 9H- purine -9- base) propane -2- base) oxygen) methyl) phosphate fumarate, structural formula is as follows:
On the other hand, the present invention relates to compound F, its chemical name is: (R)-aminomethyl phenyl ((((R) -1- (6- amino - 9H- purine -9- base) propane -2- base) oxygen) methyl) phosphate fumarate, structural formula is as follows:
On the other hand, the present invention relates to compound G, its chemical name is: (S)-ethylphenyl ((((R) -1- (6- amino - 9H- purine -9- base) propane -2- base) oxygen) methyl) phosphate fumarate, structural formula is as follows:
On the other hand, the present invention relates to compound H, its chemical name is: (R)-ethylphenyl ((((R) -1- (6- amino - 9H- purine -9- base) propane -2- base) oxygen) methyl) phosphate fumarate, structural formula is as follows:
On the other hand, the present invention relates to compound I, its chemical name is: diisopropyl 2- ((((((R) -1- (6- amino - 9H- purine -9- base) propane -2- base) oxygen) methyl) (phenoxy group) phosphinyl) amino) succinate fumarate, structural formula It is as follows:
In specific embodiments, above compound A~I is unpack format, preferably substantially pure form, more preferably With greater than about 95% purity.
On the other hand, the present invention relates to the one or more of above compound A~I to draw phenol amine rich in detection tenofovir Chinese mugwort Application in horse hydrochlorate or the sample purity for drawing the pharmaceutical preparation of phenol amine fumarate that ends containing tenofovir as reference substance, Middle tenofovir Chinese mugwort draws the chemical name of phenol amine fumarate are as follows: 9- [(R) -2- [[(R, S)-[[(S) -1- (isopropoxy carbonyl Base) ethyl] amino] phenoxy group phosphinyl] methoxyl group] propyl] adenine hemifumarate, structural formula are as follows:
Detailed description of the invention
Fig. 1: compound A:(S)-isopropyl 2- (((R)-((((R) -1- (6- amino -9H-9- purine radicals) -2- propyl) Oxygen) methyl) (methoxyl group) phosphinyl) amino) propionic ester fumarate1H-NMR；
Fig. 2: compound A:(S)-isopropyl 2- (((R)-((((R) -1- (6- amino -9H-9- purine radicals) -2- propyl) Oxygen) methyl) (methoxyl group) phosphinyl) amino) propionic ester fumarate high resolution mass spectrum；
Fig. 3: compound A:(S)-isopropyl 2- (((R)-((((R) -1- (6- amino -9H-9- purine radicals) -2- propyl) Oxygen) methyl) (methoxyl group) phosphinyl) amino) propionic ester fumarate31P-NMR；
Fig. 4: compound B:(S)-isopropyl 2- (((S)-((((R) -1- (6- amino -9H-9- purine radicals) -2- propyl) Oxygen) methyl) (methoxyl group) phosphinyl) amino) propionic ester fumarate1H-NMR；
Fig. 5: compound B:(S)-isopropyl 2- (((S)-((((R) -1- (6- amino -9H-9- purine radicals) -2- propyl) Oxygen) methyl) (methoxyl group) phosphinyl) amino) propionic ester fumarate high resolution mass spectrum；
Fig. 6: compound B:(S)-isopropyl 2- (((S)-((((R) -1- (6- amino -9H-9- purine radicals) -2- propyl) Oxygen) methyl) (methoxyl group) phosphinyl) amino) propionic ester fumarate31P-NMR；
Fig. 7: compound C:(S)-isopropyl 2- (((R)-((((R) -1- (6- amino -9H-9- purine radicals) -2- propyl) Oxygen) methyl) (ethyoxyl) phosphinyl) amino) propionic ester fumarate1H-NMR；
Fig. 8: compound C:(S)-isopropyl 2- (((R)-((((R) -1- (6- amino -9H-9- purine radicals) -2- propyl) Oxygen) methyl) (ethyoxyl) phosphinyl) amino) propionic ester fumarate high resolution mass spectrum；
Fig. 9: compound C:(S)-isopropyl 2- (((R)-((((R) -1- (6- amino -9H-9- purine radicals) -2- propyl) Oxygen) methyl) (ethyoxyl) phosphinyl) amino) propionic ester fumarate31P-NMR；
Figure 10: compound D:(S)-isopropyl 2- (((S)-((((R) -1- (6- amino -9H-9- purine radicals) -2- propyl) Oxygen) methyl) (ethyoxyl) phosphinyl) amino) propionic ester fumarate1H-NMR；
Figure 11: compound D:(S)-isopropyl 2- (((S)-((((R) -1- (6- amino -9H-9- purine radicals) -2- propyl) Oxygen) methyl) (ethyoxyl) phosphinyl) amino) propionic ester fumarate high resolution mass spectrum；
Figure 12: compound D:(S)-isopropyl 2- (((S)-((((R) -1- (6- amino -9H-9- purine radicals) -2- propyl) Oxygen) methyl) (ethyoxyl) phosphinyl) amino) propionic ester fumarate31P-NMR；
Figure 13: compound E:(S)-aminomethyl phenyl ((((R) -1- (6- amino -9H- purine -9- base) propane -2- base) oxygen) Methyl) phosphate fumarate1H-NMR；
Figure 14: compound E:(S)-aminomethyl phenyl ((((R) -1- (6- amino -9H- purine -9- base) propane -2- base) oxygen) Methyl) phosphate fumarate high resolution mass spectrum；
Figure 15: compound E:(S)-aminomethyl phenyl ((((R) -1- (6- amino -9H- purine -9- base) propane -2- base) oxygen) Methyl) phosphate fumarate31P-NMR；
Figure 16: compound F:(R)-aminomethyl phenyl ((((R) -1- (6- amino -9H- purine -9- base) propane -2- base) oxygen) Methyl) phosphate fumarate1H-NMR；
Figure 17: compound F:(R)-aminomethyl phenyl ((((R) -1- (6- amino -9H- purine -9- base) propane -2- base) oxygen) Methyl) phosphate fumarate high resolution mass spectrum；
Figure 18: compound F:(R)-aminomethyl phenyl ((((R) -1- (6- amino -9H- purine -9- base) propane -2- base) oxygen) Methyl) phosphate fumarate31P-NMR；
Figure 19: compound G:(S)-ethylphenyl ((((R) -1- (6- amino -9H- purine -9- base) propane -2- base) oxygen) Methyl) phosphate fumarate1H-NMR；
Figure 20: compound G:(S)-ethylphenyl ((((R) -1- (6- amino -9H- purine -9- base) propane -2- base) oxygen) Methyl) phosphate fumarate high resolution mass spectrum；
Figure 21: compound G:(S)-ethylphenyl ((((R) -1- (6- amino -9H- purine -9- base) propane -2- base) oxygen) Methyl) phosphate fumarate31P-NMR；
Figure 22: compound H:(R)-ethylphenyl ((((R) -1- (6- amino -9H- purine -9- base) propane -2- base) oxygen) Methyl) phosphate fumarate1H-NMR；
Figure 23: compound H:(R)-ethylphenyl ((((R) -1- (6- amino -9H- purine -9- base) propane -2- base) oxygen) Methyl) phosphate fumarate high resolution mass spectrum；
Figure 24: compound H:(R)-ethylphenyl ((((R) -1- (6- amino -9H- purine -9- base) propane -2- base) oxygen) Methyl) phosphate fumarate31P-NMR；
Figure 25: asparagus fern door propylhomoserin isopropyl ester hydrochloride1H-NMR；
Figure 26: compound I: diisopropyl 2- ((((((R) -1- (6- amino -9H- purine -9- base) propane -2- base) oxygen) Methyl) (phenoxy group) phosphinyl) amino) succinate fumarate1H-NMR；
Figure 27: compound I: diisopropyl 2- ((((((R) -1- (6- amino -9H- purine -9- base) propane -2- base) oxygen) Methyl) (phenoxy group) phosphinyl) amino) and succinate fumarate high resolution mass spectrum；
Figure 28: compound I: diisopropyl 2- ((((((R) -1- (6- amino -9H- purine -9- base) propane -2- base) oxygen) Methyl) (phenoxy group) phosphinyl) amino) succinate fumarate31P-NMR；
Figure 29: l-Alanine, N- [(S)-[[(1R) -2- (6- amino -9H-9- purine radicals) -1- methyl ethoxy] methyl] Phenoxy group phosphinyl] -, 1- Methylethyl ester, (2E) -2- butylene diester (2: 1)1H-NMR
Figure 30: l-Alanine, N- [(S)-[[(1R) -2- (6- amino -9H-9- purine radicals) -1- methyl ethoxy] methyl] Phenoxy group phosphinyl] -, 1- Methylethyl ester, the high resolution mass spectrum of (2E) -2- butylene diester (2: 1).
Figure 31: l-Alanine, N- [(S)-[[(1R) -2- (6- amino -9H-9- purine radicals) -1- methyl ethoxy] methyl] Phenoxy group phosphinyl] -, 1- Methylethyl ester, (2E) -2- butylene diester (2: 1)31P-NMR
Figure 32: impurity reference substance solution HPLC map
Figure 33: TAF test solution HPLC map
Specific embodiment
In order to which further the present invention will be described, will be illustrated below by specific embodiment, but reality below Example is applied to be not limited in any way protection scope of the present invention.
Embodiment 1:(S)-isopropyl 2- (((R, S)-((((R) -1- (6- amino -9H-9- purine radicals) -2- propyl) oxygen) Methyl) (methoxyl group) phosphinyl) amino) and propionic ester synthesis.
50ml methanol and 10ml purified water are added into the there-necked flask of 250ml, is added with stirring 5.92g9- [(R) -2- [[(R, S)-[[(S) -1- (isopropoxy carbonyl) ethyl] amino] phenoxy group phosphinyl] methoxyl group] propyl] adenine fumaric acid Salt (TAF-3), nitrogen protection are cooled to 10~20 DEG C, and 1.20g NaOH is added, finishes, is stirred overnight.Methylene chloride is added Organic phase is collected in 100ml, pure water 50ml, liquid separation, and vacuum distillation obtains solid crude product, and it is pure will to obtain crude product progress column chromatography Change, elute ratio are as follows: methanol: methylene chloride=1: 20, collect the total 230ml of eluent, 30~40 DEG C of temperature control, vacuum degree :- 0.08MPa, vacuum distillation, is evaporated off solvent to no fraction and steams, obtain yellow oil 0.67g, as (S)-isopropyl 2- (((R, S)-((((R) -1- (6- amino -9H-9- purine radicals) -2- propyl) oxygen) methyl) (methoxyl group) phosphinyl) amino) propionic ester, it receives Rate: 16.16%.
Embodiment 2:(S)-isopropyl 2- (((R, S)-((((R) -1- (6- amino -9H-9- purine radicals) -2- propyl) oxygen) Methyl) (methoxyl group) phosphinyl) amino) and propionic ester separation
By 0.67g (S)-isopropyl 2- (((R, S)-((((R) -1- (6- amino -9H-9- purine radicals) -2- propyl) oxygen) first Base) (methoxyl group) phosphinyl) amino) propionic ester is added in 50ml ethyl alcohol and dissolves, with supercritical fluid chromatography (model: MD- 2018Plus) separate sample.Column model: DAICEL CHIRALCEL AS-H (0.46cm I.D.x 25cm L), column temperature 35 DEG C, Detection wavelength: 220nm, sample introduction concentration: 20mg/ml, CO2/ EtOH=70/30, flow velocity 2.0ml/min, time 10 divide Clock.Merge collection liquid after separation, 30~40 DEG C of temperature control, vacuum degree: -0.08MPa, vacuum distillation are evaporated off solvent to no fraction and steam Out, 300.3mg (S)-isopropyl 2- (((S)-((((R) -1- (6- amino -9H-9- purine radicals) -2- propyl) oxygen) methyl) is obtained (methoxyl group) phosphinyl) amino) propionic ester (compound B free alkali), oily liquids, yield 44.78% obtains 12.2mg (S)- Isopropyl 2- (((R)-((((R) -1- (6- amino -9H-9- purine radicals) -2- propyl) oxygen) methyl) (methoxyl group) phosphinyl) ammonia Base) propionic ester (compound A free alkali), oily liquids, yield 1.82%.
Embodiment 3:(S)-isopropyl 2- (((R)-((((R) -1- (6- amino -9H-9- purine radicals) -2- propyl) oxygen) first Base) (methoxyl group) phosphinyl) amino) and propionic ester fumarate (compound A) synthesis.
10ml acetonitrile is added into the there-necked flask of 50ml, 300.3mg (S)-isopropyl 2- (((R)-is sequentially added under stirring ((((R) -1- (6- amino -9H-9- purine radicals) -2- propyl) oxygen) methyl) (methoxyl group) phosphinyl) amino) propionic ester, 84.1mg fumaric acid, nitrogen protection are heated to flowing back, and insulated and stirred 2 hours.30~40 DEG C of temperature control, vacuum degree: -0.08MPa, Vacuum distillation, is evaporated off solvent to no fraction and steams, obtain compound A, 345.7mg white powder, yield: 89.94%.1H-NMR (400Mz, DMSO-d6) δ: 13.146 (br, s, 2H), 8.141 (s, 1H), 8.097 (s, 1H), 7.201 (br, s, 2H), 6.638 (s, 2H), 5.092 (dd, J=10.4Hz, J=21.6Hz, 1H), 4.890~4.874 (m, 1H), 4.249~4.184 (m, 2H), 3.949~3.908 (m, 1H), 3.783~3.728 (m, 2H), 3.672~3.645 (m, 1H), 3.461 (d, J= 10.8Hz, 3H), 1.209 (d, J=7.2Hz, 3H), 1.170 (d, J=6.0Hz, 6H), 1.051 (d, J=6.0Hz, 3H).(in detail See attached drawing 1) ESI-HRMS spectrogram display molecular ion peak m/z=415.18585 [M+H]+, corresponding molecular weight and offer Structural formula calculated value (415.17805) is consistent.Absolute error is 1.26ppm, within high resolution mass spectrum error range. (being detailed in attached drawing 2)；31P-NMR (162Mz, DMSO-d6) δ: 25.502 (being detailed in attached drawing 3).
Embodiment 4:(S)-isopropyl 2- (((S)-((((R) -1- (6- amino -9H-9- purine radicals) -2- propyl) oxygen) first Base) (methoxyl group) phosphinyl) amino) and propionic ester fumarate (compound B) synthesis.
2ml acetonitrile is added into the there-necked flask of 25ml, 12.2mg (S)-isopropyl 2- (((S)-is sequentially added under stirring ((((R) -1- (6- amino -9H-9- purine radicals) -2- propyl) oxygen) methyl) (methoxyl group) phosphinyl) amino) propionic ester, 3.4mg Fumaric acid, nitrogen protection are heated to flowing back, and insulated and stirred 2 hours.30~40 DEG C of temperature control, vacuum degree: -0.08MPa, decompression are steamed It evaporates, solvent to no fraction is evaporated off and steams, obtains impurity B, 12.8mg white powder, yield: 81.96%.1H-NMR (400Mz, DMSO-d6) δ: 8.141 (br, s, 1H), 8.091 (br, s, 1H), 7.196 (br, s, 2H), 6.619 (s, 4H), 5.118 (dd, J =10.4Hz, J=22.0Hz, 1H), 4.911~4.880 (m, 1H), 4.275~4.110 (m, 2H), 3.907 (m, 1H), 3.794~3.700 (m, 1H), 3.688~3.639 (m, 2H), 3.459 (d, J=11.2Hz, 3H), 1.244~1.187 (m, 9H), 1.042 (d, J=6.0Hz, 3H).(being detailed in attached drawing 4) ESI-HRMS spectrogram shows molecular ion peak m/z=415.18579 [M+H]+, corresponding molecular weight is consistent with the structural formula calculated value (415.17805) of offer.Absolute error is 1.11ppm, within high resolution mass spectrum error range.(being detailed in attached drawing 5)31P-NMR (162Mz, DMSO-d6) δ: 25.116 (in detail See attached drawing 6).
Embodiment 5:(S)-isopropyl 2- (((R, S)-((((R) -1- (6- amino -9H-9- purine radicals) -2- propyl) oxygen) Methyl) (ethyoxyl) phosphinyl) amino) and propionic ester synthesis.
50ml ethyl alcohol and 10ml purified water are added into the there-necked flask of 250ml, is added with stirring 5.92g 9- [(R) -2- [[(R, S)-[[(S) -1- (isopropoxy carbonyl) ethyl] amino] phenoxy group phosphinyl] methoxyl group] propyl] adenine fumaric acid Salt (TAF-3), nitrogen protection are cooled to 10~20 DEG C, and 1.20g NaOH is added, finishes, is stirred overnight.Methylene chloride is added Organic phase is collected in 100ml, pure water 50ml, liquid separation, and vacuum distillation obtains solid crude product, and it is pure will to obtain crude product progress column chromatography Change, elute ratio are as follows: methanol: methylene chloride=1: 20, collect the total 290ml of eluent, 30~40 DEG C of temperature control, vacuum degree :- 0.08MPa, vacuum distillation, is evaporated off solvent to no fraction and steams, obtain yellow oil 1.06g, as (S)-isopropyl 2- (((R, S)-((((R) -1- (6- amino -9H-9- purine radicals) -2- propyl) oxygen) methyl) (ethyoxyl) phosphinyl) amino) propionic ester, it receives Rate: 19.91%.
Embodiment 6:(S)-isopropyl 2- (((R, S)-((((R) -1- (6- amino -9H-9- purine radicals) -2- propyl) oxygen) Methyl) (ethyoxyl) phosphinyl) amino) and propionic ester separation
By 1.00g (S)-isopropyl 2- (((R, S)-((((R) -1- (6- amino -9H-9- purine radicals) -2- propyl) oxygen) first Base) (ethyoxyl) phosphinyl) amino) propionic ester is added in 50ml ethyl alcohol and dissolves, with supercritical fluid chromatography (model: MD- 2018Plus) separate sample.Column model: DAICEL CHIRALCEL AS-H (0.46cm I.D.x 25cm L), column temperature 35 DEG C, Detection wavelength: 220nm, sample introduction concentration: 20mg/ml, CO2/ EtOH=70/30, flow velocity 2.0ml/min, time 10 divide Clock.Merge collection liquid after separation, 30~40 DEG C of temperature control, vacuum degree: -0.08MPa, vacuum distillation are evaporated off solvent to no fraction and steam Out, 468.0mg (S)-isopropyl 2- (((R)-((((R) -1- (6- amino -9H-9- purine radicals) -2- propyl) oxygen) methyl) is obtained (ethyoxyl) phosphinyl) amino) propionic ester, oily liquids, yield 46.80%.Obtain 90.0mg (S)-isopropyl 2- (((S)- ((((R) -1- (6- amino -9H-9- purine radicals) -2- propyl) oxygen) methyl) (ethyoxyl) phosphinyl) amino) propionic ester, oily Liquid, yield 9.00%.
Embodiment 7:(S)-isopropyl 2- (((R)-((((R) -1- (6- amino -9H-9- purine radicals) -2- propyl) oxygen) first Base) (ethyoxyl) phosphinyl) amino) and propionic ester fumarate (compound C) synthesis.
10ml acetonitrile is added into the there-necked flask of 50ml, 468.0mg (S)-isopropyl 2- (((R)-is sequentially added under stirring ((((R) -1- (6- amino -9H-9- purine radicals) -2- propyl) oxygen) methyl) (ethyoxyl) phosphinyl) amino) propionic ester, 128mg Fumaric acid, nitrogen protection are heated to flowing back, and insulated and stirred 2 hours.30~40 DEG C of temperature control, vacuum degree: -0.08MPa, decompression are steamed It evaporates, solvent to no fraction is evaporated off and steams, obtains compound C, 581.1mg white powder, yield: 97.70%.1H-NMR (400Mz, DMSO-d6) δ: 13.128 (br, s, 2H), 8.142 (s, 1H), 8.102 (s, 1H), 7.192 (br, s, 2H), 6.638 (s, 2H), 5.033 (dd, J=10.4Hz, J=21.6Hz, 1H), 4.891~4.860 (m, 1H), 4.292~4.149 (m, 2H), 3.940~3.713 (m, 5H), 3.654~3.631 (m, 1H), 1.225~1.138 (m, 12H), 1.055 (d, J= 6.4Hz, 3H).(being detailed in attached drawing 7) ESI-HRMS spectrogram shows molecular ion peak m/z=429.20253 [M+H]+, corresponding Molecular weight is consistent with the structural formula calculated value (429.19370) of offer.Absolute error is 3.61ppm, in high resolution mass spectrum Within error range.(being detailed in attached drawing 8)31P-NMR (162Mz, DMSO-d6) δ: 24.004 (being detailed in attached drawing 9).
Embodiment 8:(S)-isopropyl 2- (((S)-((((R) -1- (6- amino -9H-9- purine radicals) -2- propyl) oxygen) first Base) (ethyoxyl) phosphinyl) amino) and propionic ester fumarate (compound D) synthesis.
2ml acetonitrile is added into the there-necked flask of 25ml, 90.0mg (S)-isopropyl 2- (((S)-is sequentially added under stirring ((((R) -1- (6- amino -9H-9- purine radicals) -2- propyl) oxygen) methyl) (ethyoxyl) phosphinyl) amino) propionic ester, 25mg Fumaric acid, nitrogen protection are heated to flowing back, and insulated and stirred 2 hours.30~40 DEG C of temperature control, vacuum degree: -0.08MPa, decompression are steamed It evaporates, solvent to no fraction is evaporated off and steams, obtains compound D, 100.0mg white powder, yield: 87.42%.1H-NMR (400Mz, DMSO-d6) δ: 13.127 (br, s, 2H), 8.142 (s, 1H), 8.099 (s, 1H), 7.194 (br, s, 2H), 6.639 (s, 2H), 5.064 (dd, J=10.4Hz, J=22.0Hz, 1H), 4.911~4.879 (m, 1H), 4.249~4.145 (m, 2H), 3.895~3.707 (m, 4H), 3.674~3.627 (m, 2H), 1.223~1.139 (m, 12H), 1.045 (d, J= 6.4Hz, 3H).(being detailed in attached drawing 10) ESI-HRMS spectrogram display molecular ion peak m/z=429.20118 [M+H]+, it is corresponding Molecular weight is consistent with the structural formula calculated value (429.19370) of offer.Absolute error is 0.46ppm, in high resolution mass spectrum Within error range.(being detailed in attached drawing 11)31P-NMR (162Mz, DMSO-d6) δ: 23.690 (being detailed in attached drawing 12).
Embodiment 9: aminomethyl phenyl ((((R) -1- (6- amino -9H- purine -9- base) propane -2- base) oxygen) methyl) phosphoric acid The synthesis of ester.
50ml methanol is added into the there-necked flask of 250ml, is added with stirring 5.00g phenyl ((((R) -1- (6- amino -9H- Purine -9- base) propane -2- base) oxygen) methyl) phosphoryl chloride phosphorus oxychloride (TAF-2), nitrogen protection is cooled to 10~20 DEG C, and stirring 2~3 is small When.30~40 DEG C of temperature control, vacuum degree: -0.08MPa, vacuum distillation are evaporated off solvent to no fraction and steam, obtain brown oil liquid The crude product liquid is carried out HPLC separation, obtains 0.45g aminomethyl phenyl ((((R) -1- (6- amino -9H- purine -9- base) third by body Alkane -2- base) oxygen) methyl) phosphate crude product, yield: 9.11%.
Embodiment 10: aminomethyl phenyl ((((R) -1- (6- amino -9H- purine -9- base) propane -2- base) oxygen) methyl) phosphoric acid The separation of ester crude product；
By 0.45g aminomethyl phenyl ((((R) -1- (6- amino -9H- purine -9- base) propane -2- base) oxygen) methyl) phosphate Crude product, which is added in 50ml methanol, to be dissolved, and separates sample with supercritical fluid chromatography (model: MD-2018Plus).Chromatography column type Number: DAICEL CHIRALCEL AS-H (0.46cm I.D.x 25cm L), 35 DEG C of column temperature, Detection wavelength: 220nm, sample introduction are dense Degree: 20mg/ml, CO2/ MeOH=70/30, flow velocity 2.0ml/min, the time 10 minutes.Merge collection liquid after separation, temperature control 30~ 40 DEG C, vacuum degree: -0.08MPa, vacuum distillation are evaporated off solvent to no fraction and steam, obtain 110.0mg (R)-aminomethyl phenyl ((((R) -1- (6- amino -9H- purine -9- base) propane -2- base) oxygen) methyl) phosphate, oily liquids, yield: 24.44%； Obtain 316.2mg (S)-aminomethyl phenyl ((((R) -1- (6- amino -9H- purine -9- base) propane -2- base) oxygen) methyl) phosphoric acid Ester, oily liquids, yield: 70.26%.
Embodiment 11:(S)-aminomethyl phenyl ((((R) -1- (6- amino -9H- purine -9- base) propane -2- base) oxygen) methyl) The synthesis of phosphate fumarate (compound E).
10ml acetonitrile is added into the there-necked flask of 50ml, 316.2mg (S)-aminomethyl phenyl ((((R)-is sequentially added under stirring 1- (6- amino -9H- purine -9- base) propane -2- base) oxygen) methyl) phosphate, 88.21mg fumaric acid, nitrogen protection is heated to Reflux, insulated and stirred 2 hours.30~40 DEG C of temperature control, vacuum degree: -0.08MPa, vacuum distillation are evaporated off solvent to no fraction and steam Out, compound E, 370mg, white powder, yield: 98.85% are obtained.1H-NMR (400Mz, DMSO-d6) δ: 8.145 (s, 1H), 8.056 (s, 1H), 7.342 (t, J=8.0Hz, 2H), 7.226~7.194 (m, 3H), 7.056 (d, J=7.6Hz, 2H), 6.640 (s, 2H), 4.271~4.099 (m, 2H), 4.077~3.964 (m, 3H), 3.729 (d, J=10.8Hz, 3H), 1.135 (d, J=6.4Hz, 3H).(being detailed in attached drawing 13) ESI-HRMS spectrogram display molecular ion peak m/z=378.13339 [M+H]+, institute Corresponding molecular weight is consistent with the structural formula calculated value (378.12529) of offer.Absolute error is 2.18ppm, in high score It distinguishes within mass spectrum error range.(being detailed in attached drawing 14)31P-NMR (162Mz, DMSO-d6) δ: 19.886 (being detailed in attached drawing 15).
Embodiment 12:(R)-aminomethyl phenyl ((((R) -1- (6- amino -9H- purine -9- base) propane -2- base) oxygen) methyl) The synthesis of phosphate fumarate (compound F).
10ml acetonitrile is added into the there-necked flask of 50ml, is sequentially added under stirring, 110.0mg (R)-aminomethyl phenyl ((((R) -1- (6- amino -9H- purine -9- base) propane -2- base) oxygen) methyl) phosphate, 33.8mg fumaric acid, nitrogen protection, It is heated to flowing back, insulated and stirred 2 hours.30~40 DEG C of temperature control, vacuum degree: -0.08MPa, vacuum distillation are evaporated off solvent to nothing and evaporate Divide and steam, obtains compound F, 140.4mg white powder, yield: 97.61%.1H-NMR (400Mz, DMSO-d6) δ: 8.150 (s, 1H), 8.058 (s, 1H), 7.372 (t, J=8.0Hz, 2H), 7.229~7.189 (m, 3H), 7.127 (d, J=8.4Hz, 2H), 6.640 (s, 2H), 4.274~4.167 (m, 2H), 4.062~4.003 (m, 3H), 3.675 (d, J=10.8Hz, 3H), 1.113 (d, J=6.0Hz, 3H).(being detailed in attached drawing 16) ESI-HRMS spectrogram shows molecular ion peak m/z=378.13294 [M+ H]+, corresponding molecular weight is consistent with the structural formula calculated value (378.12529) of offer.Absolute error is 1ppm, in height Within Resolution Mass Spectrometry error range.(being detailed in attached drawing 17)31P-NMR (162Mz, DMSO-d6) δ: 20.019 (being detailed in attached drawing 18).
Embodiment 13: ethylphenyl ((((R) -1- (6- amino -9H- purine -9- base) propane -2- base) oxygen) methyl) phosphoric acid The synthesis of ester.
50ml ethyl alcohol is added into the there-necked flask of 250ml, is added with stirring 5.00g phenyl ((((R) -1- (6- amino -9H- Purine -9- base) propane -2- base) oxygen) methyl) phosphoryl chloride phosphorus oxychloride (TAF-2), nitrogen protection is cooled to 10~20 DEG C, and stirring 2~3 is small When.30~40 DEG C of temperature control, vacuum degree: -0.08MPa, vacuum distillation are evaporated off solvent to no fraction and steam, obtain brown oil liquid The crude product liquid is carried out HPLC separation, obtains 0.69g ethylphenyl ((((R) -1- (6- amino -9H- purine -9- base) third by body Alkane -2- base) oxygen) methyl) phosphate crude product, yield: 13.46%.
Embodiment 14: ethylphenyl ((((R) -1- (6- amino -9H- purine -9- base) propane -2- base) oxygen) methyl) phosphoric acid The separation of ester crude product；
By 0.69g ethylphenyl ((((R) -1- (6- amino -9H- purine -9- base) propane -2- base) oxygen) methyl) phosphate Crude product, which is added in 50ml methanol, to be dissolved, and separates sample with supercritical fluid chromatography (model: MD-2018Plus).Chromatography column type Number: DAICEL CHIRALCEL AS-H (0.46cm I.D.x 25cm L), 35 DEG C of column temperature, Detection wavelength: 220nm, sample introduction are dense Degree: 20mg/ml, CO2/ MeOH=70/30, flow velocity 2.0ml/min, the time 10 minutes.Merge collection liquid after separation, temperature control 30~ 40 DEG C, vacuum degree: -0.08MPa, vacuum distillation are evaporated off solvent to no fraction and steam, obtain 144.6mg (R)-ethylphenyl ((((R) -1- (6- amino -9H- purine -9- base) propane -2- base) oxygen) methyl) phosphate, oily liquids, yield: 20.96%； Obtain 305.1mg (S)-ethylphenyl ((((R) -1- (6- amino -9H- purine -9- base) propane -2- base) oxygen) methyl) phosphoric acid Ester, oily liquids, yield: 44.22%.
Embodiment 15:(S)-ethylphenyl ((((R) -1- (6- amino -9H- purine -9- base) propane -2- base) oxygen) methyl) The synthesis of phosphate fumarate (compound G).
10ml acetonitrile is added into the there-necked flask of 50ml, 305.1mg (S)-ethylphenyl ((((R)-is sequentially added under stirring 1- (6- amino -9H- purine -9- base) propane -2- base) oxygen) methyl) phosphate, 42.9mg fumaric acid, nitrogen protection is heated to Reflux, insulated and stirred 2 hours.30~40 DEG C of temperature control, vacuum degree: -0.08MPa, vacuum distillation are evaporated off solvent to no fraction and steam Out, compound G, 389.4mg white powder, yield: 98.32% are obtained.1H-NMR (400Mz, DMSO-d6) δ: 13.162 (br, 1H), 8.141 (s, 1H), 8.055 (s, 1H), 7.345 (t, J=8.0Hz, 2H), 7.219~7.174 (m, 3H), 7.061 (d, J =8.8Hz, 2H), 6.637 (s, 2H), 4.260~4.091 (m, 2H), 4.074~3.939 (m, 5H), 1.203~1.168 (m, 3H), 1.134 (d, J=6.4Hz, 3H).(being detailed in attached drawing 19) ESI-HRMS spectrogram shows molecular ion peak m/z=392.14867 [M+H]+, corresponding molecular weight is consistent with the structural formula calculated value (392.14094) of offer.Absolute error is 1.16ppm, within high resolution mass spectrum error range.(being detailed in attached drawing 20)31P-NMR (162Mz, DMSO-d6) δ: 18.558 (in detail See attached drawing 21).
Embodiment 16:(R)-ethylphenyl ((((R) -1- (6- amino -9H- purine -9- base) propane -2- base) oxygen) methyl) The synthesis of phosphate fumarate (compound H).
10ml acetonitrile is added into the there-necked flask of 50ml, 144.6mg (R)-ethylphenyl ((((R)-is sequentially added under stirring 1- (6- amino -9H- purine -9- base) propane -2- base) oxygen) methyl) phosphate, 42.9mg fumaric acid, nitrogen protection is heated to Reflux, insulated and stirred 2 hours.30~40 DEG C of temperature control, vacuum degree: -0.08MPa, vacuum distillation are evaporated off solvent to no fraction and steam Out, impurity H, 183.3mg, white powder, yield: 97.76% are obtained.1H-NMR (400Mz, DMSO-d6) δ: 13.139 (br, 1H), 8.145 (s, 1H), 8.057 (s, 1H), 7.375 (t, J=8.0Hz, 2H), 7.226~7.191 (m, 3H), 7.134 (d, J =8.8Hz, 2H), 6.635 (s, 2H), 4.273~4.200 (m, 2H), 4.043~3.986 (m, 5H), 1.168~1.099 (m, 6H).(being detailed in attached drawing 22) ESI-HRMS spectrogram display molecular ion peak m/z=392.14863 [M+H]+, corresponding molecular weight It is consistent with the structural formula calculated value (392.14094) of offer.Absolute error is 1.04ppm, in high resolution mass spectrum error model Within enclosing.(being detailed in attached drawing 23)31P-NMR (162Mz, DMSO-d6) δ: 18.654 (being detailed in attached drawing 24).
Embodiment 17: the synthesis of asparagus fern door propylhomoserin isopropyl ester hydrochloride
100ml isopropanol is added into the there-necked flask of 250ml, opens stirring, 10.00g asparagus fern door propylhomoserin is added, adds 89.42g thionyl chloride, is warming up to reflux, and insulated and stirred 6~8 hours.Vacuum distillation, 30~40 DEG C of temperature control, vacuum degree :- 0.08MPa is evaporated off solvent to no fraction and steams, and the mashing of 100ml methyl tertiary butyl ether(MTBE), filtering, by filter cake are added into residue It is dried under reduced pressure in case, is dried under reduced pressure 2~3 hours as 50 DEG C, obtain 18.49g asparagus fern door propylhomoserin salt isopropyl ester hydrochlorate, canescence Powder, yield: 97.31%.1H-NMR (400Mz, DMSO-d6) δ: 8.706 (br, 2H), 5.018~4.909 (m, 2H), 4.229 (t, J=6.0Hz, 1H), 3.004~2.895 (m, 2H), 1.238~1.192 (m, 12H) (being detailed in attached drawing 25).
Embodiment 18: diisopropyl 2- ((((((R) -1- (6- amino -9H- purine -9- base) propane -2- base) oxygen) methyl) (phenoxy group) phosphinyl) amino) succinate fumarate (compound I) synthesis
100ml methylene chloride is added into the there-necked flask of 250ml, opens stirring, sequentially adds 16.57g asparagus fern door propylhomoserin Isopropyl ester hydrochloride and 13.11g saleratus are stirred at room temperature 3~4 hours, and filtering collects filtrate, is transferred to 250ml there-necked flask In, nitrogen protection, 5.00g phenyl ((((R) -1- (6- amino -9H- purine-is slowly added dropwise into system for temperature control -10~-20 DEG C 9- yl) propane -2- base) oxygen) methyl) phosphoryl chloride phosphorus oxychloride (TAF-2) toluene (50ml) solution, drop finish, be stirred overnight.It is pure that 50ml is added Change water, extract liquid separation, collects organic phase, 30~40 DEG C of temperature control, vacuum degree: -0.08MPa, vacuum distillation are evaporated off solvent to nothing and evaporate Divide and steam, obtains 4.62g brownish black oily liquids.It takes the 1.00g oily liquids to carry out HPLC separation, obtains 0.55g diisopropyl Base 2- ((((((R) -1- (6- amino -9H- purine -9- base) propane -2- base) oxygen) methyl) (phenoxy group) phosphinyl) amino) amber Amber acid esters, pale yellow oily liquid, yield 34.48%.
10mL acetonitrile is added into the there-necked flask of 50ml, opens stirring, sequentially adds 0.55g diisopropyl 2- ((((((R) -1- (6- amino -9H- purine -9- base) propane -2- base) oxygen) methyl) (phenoxy group) phosphinyl) amino) succinic acid Ester and 0.11g fumaric acid, are heated to flowing back, and insulated and stirred 2 hours.Vacuum distillation, 30~40 DEG C of temperature control, vacuum degree :- 0.08MPa is evaporated off solvent to no fraction and steams to get compound I, 0.64g, off-white powder, yield: 96.55% is arrived.1H- NMR (400Mz, DMSO-d6) δ: 13.098 (br, 1H) 8.143 (s, 1H), 8.100 (s, 1H), 7.296 (t, J=7.6Hz, 2H), 7.161~7.143 (m, 3H), 7.080 (d, J=7.6Hz, 2H), 6.634 (s, 2H), 5.689 (m, 1H), 4.867~ 4.831 (m, 2H), 4.339~4.169 (m, 3H), 3.795~3.761 (m, 5H), 1.176~1.062 (m, 15H).It (is detailed in attached Figure 26) ESI-HRMS spectrogram show molecular ion peak m/z=563.24049 [M+H]+, the knot of corresponding molecular weight and offer Structure formula calculated value (563.23048) is consistent.Absolute error is 4.86ppm, within high resolution mass spectrum error range.(in detail See attached drawing 27)31P-NMR (162Mz, DMSO-d6) δ: 22.674,22.601 (being detailed in attached drawing 28).
Embodiment 19:((((R) -1- (6- amino -9H- purine -9- base) propane -2- base) oxygroup) methyl) phosphoric acid monophenyl (TAF-1)
Under nitrogen protection, into the reaction flask of 5L, 950g acetonitrile is added, opens stirring, sequentially adds 150g tenofovir (SM1), 106g triethylamine, 96g 4-dimethylaminopyridine and 90 grams of triphenyl phosphites (SM3).Charging finishes, and is warming up to 60 DEG C, insulated and stirred to TLC detection raw material SM1 fundamental reaction finishes.The lower vacuum distillation of stirring, is added 400g methanol to residue. 20~25 DEG C of temperature control, under stirring condition, 150g concentrated hydrochloric acid is added dropwise, time for adding 0.5~1 hour, is added dropwise, stirring 0.5 is small It is precipitated up to a large amount of solids, 2.0Kg ethyl acetate is added, stirred 12~16 hours, filtering, under the conditions of 60 DEG C, forced air drying 4 ~6 hours, obtain intermediate TAF-1 crude product about 125g.
750g water is added into 1L reaction flask, opens stirring, adds 125g TAF-1 crude product, 10~20 DEG C of temperature control, stir Mix 0.5~1 hour, filter, with 237g ethanol rinse, under the conditions of 55~65 DEG C, forced air drying 6~8 hours, obtain for class it is white Color powder 102g.Yield: 55.0%.
Embodiment 20:9- [(R) -2- [[(R, S)-[[(S) -1- (isopropoxy carbonyl) ethyl] amino] phenoxy group phosphine oxide Base] methoxyl group] propyl] adenine (TAF-3)
600g toluene is added to the reaction flask of 5L, opens stirring, sequentially adds 100g TAF-1 and 300g thionyl chloride. It 70 DEG C of temperature control, stirs 24 hours.After reaction, it is evaporated under reduced pressure, 600g normal heptane is added to residue, stirring 0.5~1 is small When, filtering, filter cake is eluted with 300g normal heptane, and forced air drying 8~12 hours under the conditions of 40 DEG C obtain TAF-2 94g, for Huang Color powder.
1.8Kg methylene chloride is added into 5L reaction flask, opens stirring, sequentially adds 180g l-Alanine isopropyl ester (SM2), with 60g eluent methylene chloride, mother liquor is transferred in 5L reaction flask with 400g saleratus, stirring 2 hours, filtering, Stirring is opened, 180g anhydrous sodium sulfate is added, is stirred 0.5~1 hour, with 60g eluent methylene chloride, mother liquor is shifted for filtering Into 5L reaction flask, under nitrogen gas stirring, opens stirring and be added dropwise under conditions of control reaction temperature is 0~10 DEG C into system The toluene mixed serum (90g TAF-2 being added in 610g toluene, stir 1 hour for use) of 700g TAF-2, time for adding 1~2 Hour, it is added dropwise, continues insulated and stirred 1~2 hour.900g water, stirring are added into reaction system, liquid separation is collected organic Phase, then successively purify water washing with 15% sodium dihydrogen phosphate of 450g (W/W) aqueous solution and 900g respectively, collect organic phase.To have Machine phase transfer opens stirring into 5L reaction flask, and 100g anhydrous sodium sulfate is added, and stirs 0.5~1 hour, filtering, with 60g bis- Chloromethanes elution.Mother liquor is transferred in 5L reaction flask, is evaporated under reduced pressure.
1100g acetonitrile is added to residue, opens stirring, stirs 2~3 hours under the conditions of 40 DEG C of temperature control, filter while hot, receive Collect filtrate.40 DEG C of temperature control, stirring continues stirring 0.5 hour to system dissolved clarification, and slow cooling is to 0~5 DEG C, temperature fall time 2~3 Hour, insulated and stirred 2~3 hours.Filtering, forced air drying 12 hours under the conditions of 40 DEG C obtain off-white powder (TAF-3) 53.8g, yield: 41.1%.
Embodiment 21:9- [(R) -2- [[(S)-[[(S) -1- (isopropoxy carbonyl) ethyl] amino] phenoxy group phosphinyl] Methoxyl group] propyl] adenine hemifumarate (TAF)
Under nitrogen protection, 200g isopropanol, 9 grams of fumaric acid, 50 grams of TAF-3 are sequentially added into 2L reaction flask.Unlatching is stirred It mixes, under the conditions of 40~50 DEG C of temperature control, stirs dissolved clarification, filter while hot, collect mother liquor.Mother liquor is transferred in 2L reaction flask, is opened Stirring under the conditions of 40~50 DEG C of temperature control, stirs dissolved clarification, and slow cooling is to 0~5 DEG C, temperature fall time 2~3 hours, insulated and stirred 10 Hour.Filtering forced air drying 12~16 hours, obtains tenofovir Chinese mugwort and draws phenol amine hemifumarate under the conditions of 60~65 DEG C Finished product 37.5g is white powder, yield 75.0%.1H-NMR (400Mz, DMSO-d6) δ: 13.140 (br, 1H), 8.158 (s, 1H), 8.117 (s, 1H), 7.303 (t, J=7.6Hz, 2H), 7.303 (t, J=7.6Hz, 2H), 7.123 (t, J=7.2Hz, 1H), 7.077 (d, J=7.6Hz, 2H), 6.648 (s, 1H), 5.613 (t, J=12.0Hz, 1H), 4.860 (m, 1H), 4.276 (m, 1H), 4.153 (dd, J=6.4,14.4Hz, 1H), 3.948 (m, 1H), 3.856~3.909 (m, 2H), 3.77 (dd, J= 9.6,13.6Hz, 1H), 1.155~1.170 (m, 6H), 1.141~1.155 (m, 3H), 1.058 (d, J=6.4Hz, 3H).(in detail See attached drawing 29) ESI-HRMS spectrogram display molecular ion peak m/z=477.20187 [M+H]+, corresponding molecular weight and offer Structural formula calculated value (477.20098) be consistent.Absolute error is 1.87ppm, within high resolution mass spectrum error range. (being detailed in attached drawing 30)31P-NMR (162Mz, DMSO-d6) δ: 22.120 (being detailed in attached drawing 31).
Embodiment 22: impurity A-I ends in tenofovir as impurity reference substance and draws phenol amine hemifumarate bulk pharmaceutical chemicals impurity Application in detection
(1) preparation of impurity A-I reference substance solution:
Take the impurity A-I of above-mentioned preparation appropriate, it is accurately weighed, add appropriate 80% acetonitrile-water ultrasound to make to dissolve, with 80% second Nitrile-water quantitatively dilute be made in every 1ml be respectively containing about compound A~I 1 μ g mixed solution, mixed as impurity reference substance Solution.
(2) preparation of test solution:
The tenofovir Chinese mugwort of above-mentioned preparation draws the preparation of the test solution of phenol amine hemifumarate: taking according to above-mentioned implementation Tenofovir Chinese mugwort prepared by the step of example 19~21 draws phenol amine hemifumarate appropriate (Chinese mugwort containing tenofovir draws phenol amine about 10mg), It is accurately weighed, it sets in 10ml volumetric flask, adds 80% acetonitrile-water to dissolve and be diluted to scale, shake up, every 1ml is made and contains for promise good fortune The solution of Wei Aila phenol amine about 1mg, as test solution.
(3) chromatographic condition is as follows:
It is filler with octadecylsilane chemically bonded silica chromatographic column (4.6 × 150mm, 2.7 μm), with water (0.1% first Acid) it is mobile phase A, with methanol (contain 0.1% formic acid) for Mobile phase B, gradient elution is carried out according to following table, flow velocity 1.0ml/min, Detection wavelength 260nm, 40 DEG C of column temperature, 10 μ l of sampling volume.
Gradient elution program:
Time (minute) Mobile phase A (%) Mobile phase B (%)
0 95 5
10 85 15
40 55 45
50 10 90
55 10 90
55.01 95 5
60 95 5
(4) test procedure: 10 μ l of above-mentioned impurity reference substance solution and test solution is taken, liquid chromatograph is injected separately into (being purchased from U.S. Agilent company, model: 1260 HPLC of Agilent) or LC-MS instrument (Agilent company, the U.S., Agilent 6120), chromatogram is recorded, calculates each impurity A-according to external standard method (four general rules 0512 of Chinese Pharmacopoeia version in 2015) The content of I (referring to Figure 32,33).
(5) test result:
The tenofovir Chinese mugwort prepared according to embodiment 19~21 draws the impurity spectrum analysis of phenol amine hemifumarate
It should be noted that all documents referred in the present invention are incorporated as referring in this application, just as each Piece document is individually recited as with reference to such.In addition, it should also be understood that, above-described is that specific implementation of the invention is arranged and transported Technical principle, after having read the contents of the present invention, those skilled in the art can do various changes or be repaired to the present invention Change without departing from spirit and scope of the invention, such equivalent forms are also fallen in the scope of the present invention.

Claims (11)

1. compound A, its chemical name is: (S)-isopropyl 2- (((R)-((((R) -1- (6- amino -9H-9- purine radicals) -2- Propyl) oxygen) methyl) (methoxyl group) phosphinyl) amino) propionic ester fumarate (17-A), structural formula is as follows:
2. compound B, its chemical name is: (S)-isopropyl 2- (((S)-((((R) -1- (6- amino -9H-9- purine radicals) -2- Propyl) oxygen) methyl) (methoxyl group) phosphinyl) amino) propionic ester fumarate (17-B), structural formula is as follows:
3. compound C, its chemical name is: (S)-isopropyl 2- (((R)-((((R) -1- (6- amino -9H-9- purine radicals) -2- Propyl) oxygen) methyl) (ethyoxyl) phosphinyl) amino) propionic ester fumarate (19-A), structural formula is as follows:
4. compound D, its chemical name is: (S)-isopropyl 2- (((S)-((((R) -1- (6- amino -9H-9- purine radicals) -2- Propyl) oxygen) methyl) (ethyoxyl) phosphinyl) amino) propionic ester fumarate (19-B), structural formula is as follows:
5. compound E, its chemical name is: (S)-aminomethyl phenyl ((((R) -1- (6- amino -9H- purine -9- base) propane -2- Base) oxygen) methyl) phosphate fumarate (18-A), structural formula is as follows:
6. compound F, its chemical name is: (R)-aminomethyl phenyl ((((R) -1- (6- amino -9H- purine -9- base) propane -2- Base) oxygen) methyl) phosphate fumarate (18-B), structural formula is as follows:
7. compound G, its chemical name is: (S)-ethylphenyl ((((R) -1- (6- amino -9H- purine -9- base) propane -2- Base) oxygen) methyl) phosphate fumarate (4-A), structural formula is as follows:
8. compound H, its chemical name is: (R)-ethylphenyl ((((R) -1- (6- amino -9H- purine -9- base) propane -2- Base) oxygen) methyl) phosphate fumarate (4-B), structural formula is as follows:
9. compound I, its chemical name is: diisopropyl 2- ((((((R) -1- (6- amino -9H- purine -9- base) propane -2- Base) oxygen) methyl) (phenoxy group) phosphinyl) amino) succinate fumarate (23) its structural formula is as follows:
It is unpack format, preferably substantially pure form 10. compound described according to claim 1~any one of 9, more It is preferred that having greater than about 95% purity.
11. the one or more of compound described according to claim 1~any one of 9 draw phenol half in detection tenofovir Chinese mugwort Application in fumarate or the sample purity for drawing the pharmaceutical preparation of phenol hemifumarate that ends containing tenofovir as reference substance, Wherein tenofovir Chinese mugwort draws the chemical name of phenol hemifumarate are as follows: (9- [(R) -2- [[(S)-[[(S) -1- (isopropoxy carbonyl Base) ethyl] amino] phenoxy group phosphinyl] methoxyl group] propyl] adenine hemifumarate, structural formula are as follows:
CN201810099676.9A 2018-01-31 2018-01-31 Tenofovir Chinese mugwort draws the preparation and its application of phenol amine fumarate process impurity Pending CN110092803A (en)

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CN111018914A (en) * 2019-12-16 2020-04-17 株洲千金药业股份有限公司 Preparation method of tenofovir disoproxil fumarate impurity
CN111018916A (en) * 2019-12-30 2020-04-17 天津天士力圣特制药有限公司 Synthesis method of tenofovir phenyl alkyl ester phosphonamide precursor
CN111620908A (en) * 2020-05-20 2020-09-04 北京华睿鼎信科技有限公司 Diastereoisomer of tenofovir alafenamide, preparation method and application thereof
CN113777186A (en) * 2021-08-12 2021-12-10 北京鑫开元医药科技有限公司 Method for detecting impurities in propane fumarate tenofovir
CN114292297A (en) * 2021-12-21 2022-04-08 福建南方制药股份有限公司 Method for preparing antiviral drug tenofovir alafenamide fumarate
CN114605472A (en) * 2021-12-27 2022-06-10 石家庄龙泽制药股份有限公司 TAF oxidation degradation impurity, preparation method and application
CN116082252A (en) * 2022-09-21 2023-05-09 桂林医学院第二附属医院 Carmof hydrate, preparation method, pharmaceutical composition and application thereof

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CN111018914A (en) * 2019-12-16 2020-04-17 株洲千金药业股份有限公司 Preparation method of tenofovir disoproxil fumarate impurity
CN111018916A (en) * 2019-12-30 2020-04-17 天津天士力圣特制药有限公司 Synthesis method of tenofovir phenyl alkyl ester phosphonamide precursor
CN111620908A (en) * 2020-05-20 2020-09-04 北京华睿鼎信科技有限公司 Diastereoisomer of tenofovir alafenamide, preparation method and application thereof
CN113777186A (en) * 2021-08-12 2021-12-10 北京鑫开元医药科技有限公司 Method for detecting impurities in propane fumarate tenofovir
CN113777186B (en) * 2021-08-12 2023-06-13 北京鑫开元医药科技有限公司 Method for detecting impurities in propionofovir fumarate
CN114292297A (en) * 2021-12-21 2022-04-08 福建南方制药股份有限公司 Method for preparing antiviral drug tenofovir alafenamide fumarate
CN114292297B (en) * 2021-12-21 2024-05-10 福建南方制药股份有限公司 Method for preparing antiviral drug tenofovir alafenamide fumarate
CN114605472A (en) * 2021-12-27 2022-06-10 石家庄龙泽制药股份有限公司 TAF oxidation degradation impurity, preparation method and application
CN116082252A (en) * 2022-09-21 2023-05-09 桂林医学院第二附属医院 Carmof hydrate, preparation method, pharmaceutical composition and application thereof
CN116082252B (en) * 2022-09-21 2024-06-11 桂林医学院第二附属医院 Carmof hydrate, preparation method, pharmaceutical composition and application thereof

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