WO2011103762A1 - 拉米夫定草酸盐及其制备方法 - Google Patents

拉米夫定草酸盐及其制备方法 Download PDF

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WO2011103762A1
WO2011103762A1 PCT/CN2010/080151 CN2010080151W WO2011103762A1 WO 2011103762 A1 WO2011103762 A1 WO 2011103762A1 CN 2010080151 W CN2010080151 W CN 2010080151W WO 2011103762 A1 WO2011103762 A1 WO 2011103762A1
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lamivudine
alkyl group
oxalate
group
formula
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PCT/CN2010/080151
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English (en)
French (fr)
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刘彦龙
刘飞
张喜全
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江苏正大天晴药业股份有限公司
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Priority to KR1020127025959A priority Critical patent/KR101525296B1/ko
Priority to EP20100846385 priority patent/EP2540717B1/en
Priority to JP2012554198A priority patent/JP5613780B2/ja
Priority to US13/581,268 priority patent/US8569322B2/en
Publication of WO2011103762A1 publication Critical patent/WO2011103762A1/zh

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    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D411/00—Heterocyclic compounds containing two or more hetero rings, at least one ring having oxygen and sulfur atoms as the only ring hetero atoms
    • C07D411/02—Heterocyclic compounds containing two or more hetero rings, at least one ring having oxygen and sulfur atoms as the only ring hetero atoms containing two hetero rings
    • C07D411/04—Heterocyclic compounds containing two or more hetero rings, at least one ring having oxygen and sulfur atoms as the only ring hetero atoms containing two hetero rings directly linked by a ring-member-to-ring-member bond
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/33—Heterocyclic compounds
    • A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
    • A61K31/495—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with two or more nitrogen atoms as the only ring heteroatoms, e.g. piperazine or tetrazines
    • A61K31/505—Pyrimidines; Hydrogenated pyrimidines, e.g. trimethoprim
    • A61K31/506—Pyrimidines; Hydrogenated pyrimidines, e.g. trimethoprim not condensed and containing further heterocyclic rings
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/33—Heterocyclic compounds
    • A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
    • A61K31/495—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with two or more nitrogen atoms as the only ring heteroatoms, e.g. piperazine or tetrazines
    • A61K31/505—Pyrimidines; Hydrogenated pyrimidines, e.g. trimethoprim
    • A61K31/513—Pyrimidines; Hydrogenated pyrimidines, e.g. trimethoprim having oxo groups directly attached to the heterocyclic ring, e.g. cytosine
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P31/00—Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
    • A61P31/12—Antivirals
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P31/00—Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
    • A61P31/12—Antivirals
    • A61P31/14—Antivirals for RNA viruses
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P31/00—Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
    • A61P31/12—Antivirals
    • A61P31/14—Antivirals for RNA viruses
    • A61P31/18—Antivirals for RNA viruses for HIV
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D239/00—Heterocyclic compounds containing 1,3-diazine or hydrogenated 1,3-diazine rings
    • C07D239/02—Heterocyclic compounds containing 1,3-diazine or hydrogenated 1,3-diazine rings not condensed with other rings
    • C07D239/20—Heterocyclic compounds containing 1,3-diazine or hydrogenated 1,3-diazine rings not condensed with other rings having two double bonds between ring members or between ring members and non-ring members
    • C07D239/22—Heterocyclic compounds containing 1,3-diazine or hydrogenated 1,3-diazine rings not condensed with other rings having two double bonds between ring members or between ring members and non-ring members with hetero atoms directly attached to ring carbon atoms

Definitions

  • Lamivudine oxalate and preparation method thereof
  • the invention relates to a novel salt of lamivudine and a preparation method thereof.
  • the salt of lamivudine is an oxalate of cis lamivudine. Background technique
  • Nucleoside compounds and their analogs and derivatives are an important class of antiviral therapeutics. Many nucleoside compounds have shown good antiviral activity against retroviruses such as human immunodeficiency virus (HI V ), hepatitis B virus (HBV) and human tropic cell virus (HTLV).
  • HIV V human immunodeficiency virus
  • HBV hepatitis B virus
  • HTLV human tropic cell virus
  • An antiviral nucleoside compound, lamivudine which can be represented by Formula I, is disclosed in International Application No. PCT/GB91/00706, the disclosure of which is incorporated herein by reference.
  • nucleoside compounds and derivatives thereof contain at least two chiral centers, and It exists in the form of optical isomers. However, usually only cis isomers of such compounds show strong biological activity. Therefore, it has been a subject of research to select a stereospecific synthesis method which is versatile and suitable for industrial production of nucleoside compounds.
  • W092/20699 discloses a diastereoselective method for the preparation of optically active cis-nucleoside analogs and derivatives which are predominantly lamivudine. This method relies on the use of a particular Lewis acid. Such Lewis acids are highly reactive and unstable compounds that are expensive and have significant toxicity.
  • WO/1995/029174 optimises the method provided by W092/20699, in order to avoid the addition of a Lewis acid, selecting a suitable leaving group, reacting an oxathiolane with a substituent and a leaving group and a pyrimidine
  • the salicylate of lamivudine is disclosed as an intermediate salt of refined lamivudine.
  • the inventors of the present invention directly use a compound having a chiral preparation as a raw material to prepare lamivudine, which can ensure high stereoselective cis lamivudine, and is easy to purify and separate, and is suitable for industrial applications.
  • the inventors of the present application have studied the technology disclosed in WO/1995/029174, in particular, the salicylate of lamivudine has been further evaluated, and it has been found that the salt still has many defects:
  • Lamivudine salicylate has poor crystallinity and viscous traits.
  • the skilled person can understand that the viscous material is easy to carry impurities and is difficult to purify. As an intermediate, it requires multi-step refining to produce high-purity lamivudine, so the cost is high in the industry, and multi-step purification prolongs production. Time, reducing production efficiency.
  • the lamivudine oxalate has the following structural formula:
  • Formula II shows the salt formed by cis-lamivudine, the lamivudine-left-handed body and oxalic acid.
  • Another object of the present invention is to provide a process for the preparation of the lamivudine oxalate.
  • One method is to react lamivudine and oxalic acid in a suitable solvent, wherein suitable solvents include water, a polar solvent such as ether, tetrahydrofuran, dioxane or alcohol, wherein the alcohol is, for example, methanol or ethanol, solvent It may also be a mixture of various solvents.
  • suitable solvents include water, a polar solvent such as ether, tetrahydrofuran, dioxane or alcohol, wherein the alcohol is, for example, methanol or ethanol, solvent It may also be a mixture of various solvents.
  • R is an optionally substituted alkyl group, including.
  • the alkyl group may be a straight, branched or cyclic alkyl group, and the substituent is an alkyl group, a halogen group, a hydroxyl group, a phenyl group or the like.
  • R is a chiral reagent, which facilitates separation of the enantiomers, such as menthyl, phenyl-substituted menthyl, and further preferably L-menthyl, L-8-phenylmenthyl.
  • the above oxalic acid includes an oxalic acid derivative, which is reacted with an oxalic acid or an oxalic acid derivative to be reduced and salted, wherein the reducing agent comprises sodium borohydride or potassium borohydride.
  • the reducing agent comprises sodium borohydride or potassium borohydride.
  • the step of synthesizing lamivudine oxalate from the formula III comprises: reacting the formula III with sodium borohydride or potassium borohydride in a buffer solution to remove impurities or unreacted raw materials, and then reacting with oxalic acid or oxalic acid derivatives. The reaction gives lamivudine oxalate.
  • the solution obtained in the step (1) is added dropwise with sodium borohydride or potassium borohydride solution, and the reaction is detected to be complete;
  • step (3) The product of the step (3) is added with oxalic acid or an oxalic acid derivative, and the reaction is subjected to purification treatment to obtain lamivudine oxalate crystals.
  • the buffer solution in the above step 1 is a buffer solution required to use a reducing agent sodium borohydride or potassium borohydride, a suitable buffer solution such as a phosphate or borate buffer, such as an aqueous solution of dipotassium hydrogen phosphate, a concentration of dipotassium hydrogen phosphate solution. It is 20-50%, preferably 25-25%, and the most preferred concentration is 31%.
  • the suitable temperature is 18 ⁇ 20 °C.
  • the amount of ethanol in the step 1 means 4 to 7 times, preferably 5 to 6 times the amount of the raw material.
  • the concentration of sodium borohydride (or potassium borohydride) solution is 5 mol/L, preferably prepared by the following method: first preparing a 0.4% aqueous solution of sodium hydroxide (or potassium hydroxide), and then adding a certain amount of sodium borohydride ( Or potassium borohydride) is dissolved in the sodium hydroxide (or potassium hydroxide) solution to prepare a sodium borohydride (or potassium borohydride) solution having a concentration of 5 mol/L.
  • detection to completion of the reaction means the amount of the raw material of the formula III or the formula III after the reaction, which is ⁇ 0.5%.
  • the acid for adjusting the pH in the step 3 may, for example, be hydrochloric acid, sulfuric acid, phosphoric acid or nitric acid, preferably hydrochloric acid, for example, 6 mol/L hydrochloric acid, and the pH-adjusting base may be sodium hydroxide or potassium hydroxide.
  • the lamivudine oxalate described in the present invention is cis lamivudine (lamivudine left-handed body) a salt formed with oxalic acid.
  • the chemical name of the raw material ⁇ ⁇ in the present invention is: (21, 58) _5_ (4_amino 2 - oxo-pyrimidin-1-yl) - [1, 3] oxathiolane _2_carboxylic acid, 2S_isopropyl_5R-methyl_1R-cyclohexyl ester; the above-mentioned formula III and CME starting materials are commercially available or can be prepared according to the method disclosed in WO/1995/029174.
  • Another aspect of the invention provides a method of preparing lamivudine using lamivudine oxalate as a starting material.
  • lamivudine oxalate is easily separated from polar solvents and can be converted to lamivudine free base in a conventional manner.
  • the conventional method involves the use of a suitable base, and a suitable base can be a tertiary amine such as triethylamine.
  • lamivudine oxalate is added to ethanol, and a certain amount of triethylamine is added in an hour and a half at 40 to 45 ° C, and the mixture is stirred for 4 hours, centrifuged at room temperature, and the product is vacuum dried at 40 to 45 ° C. That is, lamivudine crystals are obtained.
  • the invention also provides a method for preparing lamivudine by using the formula 111 as a raw material, comprising the reaction of the formula 111 with sodium borohydride or potassium borohydride, and then reacting with oxalic acid or an oxalic acid derivative to obtain lamivudine oxalate. And then reacted with a base to obtain lamivudine.
  • R is an optionally substituted alkyl group, including .
  • the alkyl group may be a straight, branched or cyclic alkyl group, the substituent is an alkyl group, a halogen group, a hydroxyl group, a phenyl group or the like, and R is preferably a menthyl group or a phenyl-substituted menthyl group, and further preferably L- Menthyl, L-8-phenyl menthyl.
  • the compound of formula I is obtained by reducing the formula III and forming a salt with oxalic acid or a derivative thereof, and then releasing the corresponding base, wherein the reagent in the step of formula III to formula II comprises sodium borohydride or potassium borohydride.
  • the present invention provides a crystalline form of lamivudine oxalate which provides a crystal of lamivudine oxalate.
  • the crystal uses X-ray powder diffraction spectrum of Cu-Ko radiation, 2 ⁇ expressed in degrees, and diffraction peaks of 21.04, 22.44, 23.46, 23.78, 25.26, further at 13.94, 14.86, 21.04, 21.30, 22.44, 23.46, 23.78, 25.26, 33.36 have diffraction peaks, and further have diffraction peaks at 13.94, 14.86, 16.88, 17.66, 20.34, 21.04, 21.30, 22.44, 23.46, 23.78, 25.26, 28.30, 33.36.
  • a specific embodiment of the invention discloses an X-ray powder diffraction spectrum in a situation, see Figure 1.
  • the above-described lamivudine oxalate crystals of the present invention are anhydrous, solvent-free, or contain little or no detectable water or solvent.
  • the crystals of the present invention contain less than about 1% water or solvent. Typically, it contains less than 0.5% water or solvent.
  • the crystal of the present invention may contain less than 20% of the amorphous form.
  • the amorphous form of the lamivudine oxalate salt of the present invention generally contains less than 10%, typically less than 1%, usually less than 0.1%.
  • the lamivudine oxalate crystal provided by the present invention is a DSC endothermic transition peak at about 201. 9 ° C, and an endothermic transition is initiated at about 199 ° C.
  • An example DSC spectrum is shown in Fig. 2, and the endothermic transition The temperature is 199. 23 ° C 0. Melting point: 201. 3 - 202. 7 ° C.
  • the oxalate salt of lamivudine is a pharmaceutically acceptable salt which can be used as an antiviral agent as described in W091 17159 or as a pharmaceutical composition.
  • the lamivudine oxalate prepared by the present invention is an excellent intermediate for preparing high-purity lamivudine.
  • other organic acid salts of lamivudine such as salicylate
  • the lamivudine oxalate is a good crystal, the above phenomenon does not occur in the preparation process, the lamivudine prepared by the lamivudine oxalate can be purified to 99.5%. the above. Very favorable conditions are provided for the later preparation of high purity cis lamivudine.
  • the lamivudine oxalate provided by the invention is a salt formed by one molecule of oxalic acid and two molecules of lamivudine, and the amount of triethylamine used when removing the organic acid for preparing lamivudine is higher than that of other lamivuds.
  • the organic acid salt is greatly reduced.
  • Triethylamine is a toxic, flammable chemical agent with a strong ammonia odor. It is slightly fuming in air, and its vapor or liquid can stimulate human skin and mucous membranes. It is mainly absorbed by the body through inhalation, ingestion, percutaneous, etc.
  • triethylamine After inhalation of triethylamine vapor, the human body has a strong stimulating effect on the respiratory tract, which may cause pulmonary edema or even death. Eye and skin contact may cause chemical burns. Therefore, the reduction of triethylamine reduces the cost of industrial production on the one hand, and more importantly, reduces the use of toxic reagents, improves the safety of the production process and reduces the chemical residue of the final product.
  • the inventors focused on the comparison of lamivudine oxalate and salicylate in the case of equimolar feeding.
  • the amount of triethylamine required for the organic acid was calculated.
  • the lamivudine crystals obtained by the above method were examined by liquid chromatography according to a known scheme.
  • the apparatus and chromatographic conditions were as follows:
  • Figure 1 is an X-ray diffraction pattern of lamivudine oxalate crystals
  • DSC chart differential scanning calorimetry diagram
  • Example 1 The invention is further illustrated by the following examples, which are not intended to limit the invention in any way.
  • Example 1 The invention is further illustrated by the following examples, which are not intended to limit the invention in any way.
  • the organic phases were combined and adjusted to pH 6 to 6.5 with 6 mol/L hydrochloric acid, and the temperature was maintained at 28 to 32 °C.
  • the ethanol was distilled off, cooled to room temperature, dissolved in 280 ml of water and washed twice with 240 ml of toluene.
  • the toluene layers were combined and extracted with 70 ml of water.
  • 6 g of activated carbon was added, stirred at room temperature for 30 minutes, filtered, and the filter cake was used. Wash with 180 ml of water.
  • the DSC of the lamivudine oxalate crystal form has an endothermic transition temperature starting temperature of 199.23 ° C, a peak value of 201.87 ° C, and a heating rate of 10.00 ° C / min. Melting point: 201.3-202.7 °C. Instrument model: waters2414.
  • the melting point of lamivudine was determined to be 174-177 °C.

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Abstract

本发明涉及一种拉米夫定草酸盐及其制备方法，以及拉米夫定草酸盐的晶型。本发明提供一种拉米夫定的制备方法。

Description

说明书
拉米夫定草酸盐及其制备方法
技术领域
本发明涉及一种新的拉米夫定的盐及其制备方法。 所述拉米夫定的盐为 顺式拉米夫定的草酸盐。 背景技术
核苷类化合物和它们的类似物、 衍生物是一类重要的抗病毒治疗剂。 许 多核苷类化合物对逆转录病毒，如人体免疫缺乏病毒（HI V )、 乙肝病毒（HBV ) 和人体嗜 T细胞病毒 （ HTLV )显示出了良好的抗病毒活性。 国际申请 PCT/GB91/00706 , 公开号 W091/17159中公开了可由式 I表示 的一种抗病毒核苷类化合物一一拉米夫定。
Figure imgf000002_0001
拉米夫定
4_氨基- 1_ ( 2R -羟基曱基 3]氧硫杂环戊烷 -5S -基） -1H -嘧啶 _2 -酮 多数核苷类化合物及其衍生物含有至少两个手性中心， 并以光学异构体 的形式存在。 然而， 通常此类化合物仅顺式异构体显示出较强的生物活性。 因而， 选择一个通用、 并且适合工业化生产的核苷类化合物的立体选择合成 方法一直是人们研究的课题。 W092/20699 公开了一种用于制备主要为拉米夫定的光学活性顺式核苷 类似物和衍生物的非对映选择方法。 该方法依赖于使用特定的路易斯酸。 而 此类路易斯酸是一种高反应性的和不稳定的化合物，昂贵并且有明显的毒性。
WO/1995/029174对 W092/20699提供的方法进行了优化， 为了避免添加 路易斯酸， 选择合适的离去基团， 将带有取代基和离去基团的氧硫杂环戊烷 和嘧啶反应， 并且公开了拉米夫定的水杨酸盐， 作为精制拉米夫定的一种中 间体盐。 本申请发明人直接使用带有手性制剂的化合物作为原料， 制备拉米夫 定， 可以保证得到高立体选择性的顺式拉米夫定， 并且容易纯化分离， 适合 于工业应用。 本申请的发明人对 WO/ 1995/ 029174 所公开的技术进行了研究， 尤其是 拉米夫定的水杨酸盐作了进一步评价， 发现该盐依旧存在诸多缺陷：
1、 拉米夫定水杨酸盐结晶性较差， 性状发粘。 技术人员能够了解性状 发粘的物质容易携带杂质， 不易纯化， 以此作为中间体需要多步精制才能生 产出高纯度的拉米夫定， 因此工业上成本较高， 并且多步纯化延长了生产时 间， 降低了生产效率。
2、 WO/1995/029174 中为脱去水杨酸而大量添加三乙胺， 而三乙胺对人 体有刺激和毒副作用， 在拉米夫定的质量控制中需要严格控制， 为了保证该 化学试剂的残留符合质量标准， 需要建立一套相应的质量控制标准。 并且大 量添加三乙胺不利于工业生产中控制生产成本。 为了得到高立体选择性， 高纯度的拉米夫定， 又节约成本， 方便工业生 产， 并且减少产品的毒副作用， 本申请的发明人完成了本发明。 发明内容
本发明一个目的是提供一种拉米夫定草酸盐。 所述的拉米夫定草酸盐具 有如下结构式：
Figure imgf000004_0001
拉米夫定草酸盐
Figure imgf000004_0002
式 II所标示的是顺式拉米夫定即拉米夫定左旋体与草酸形成的盐。
本发明的另一个目的是提供所述拉米夫定草酸盐的制备方法。
一种方法为将拉米夫定和草酸在合适的溶剂中反应， 其中合适的溶剂包 括水、 极性溶剂， 例如醚、 四氢呋喃、 二噁烷或醇， 其中的醇例如为甲醇或 乙醇， 溶剂也可以是各种溶剂的混合物。
另一种制备方法步骤如下：
Figure imgf000004_0003
式 III
其中 R为任意取代的烷基， 包括 。烷基， 该烷基可以是直连、 支链或 环烷基， 取代基为烷基、 卤素、 羟基、 苯基等。 优选 R为手性试剂， 利于对 映体的分离，例如薄荷烷基、苯基取代的薄荷烷基，进一步优选 L-薄荷烷基、 L-8-苯基薄荷烷基。 上述的草酸包括草酸衍生物， 将式 I I I和草酸或草酸衍 生物反应， 还原并成盐， 其中的还原试剂包括硼氢化钠或硼氢化钾。。 可以例举的一个实例为如下:
Figure imgf000005_0001
式 ΙΙΓ
以式 III为原料合成拉米夫定草酸盐的步骤包括： 在緩沖溶液中式 III和硼 氢化钠或硼氢化钾反应， 去除杂质或未反应完全的原料后， 然后再和草酸或 草酸衍生物反应， 得到拉米夫定草酸盐。
具体步骤如下：
( 1 )、 制备緩沖溶液， 加入适量乙醇搅拌均勾， 所得溶液中加入原料式
III；
( 2 )、 将步骤（ 1 )所得溶液滴加硼氢化钠或硼氢化钾溶液， 检测至反 应完全；
( 3)、 将步骤（2 )所得溶液用溶剂或水处理， 去除杂质或未反应完全 的原料；
(4 )、 步骤（ 3)的产物加入草酸或草酸衍生物， 反应， 纯化处理， 即得 拉米夫定草酸盐晶体。
再进一步具体为：
( 1 )、制备緩沖溶液，加入适量乙醇搅拌均匀 ,所得溶液中加入原料 CME; ( 2)、 步骤（1 )所得溶液在 1 小时内滴加硼氢化钠溶液， 检测至反应 完全； ( 3)、 将步骤（2)所得溶液静置分层， 下层水层用适量乙醇洗涤， 将 所有有机溶剂层合并后用酸调 pH至 6 ~ 6.5, 温度保持在 28 ~ 32°C。再用碱 调 pH至 8.0~8.3。 将乙醇蒸掉， 降至室温， 加入适量水溶解并用有机溶剂 洗涤，有机溶剂层合并后用水萃取，合并所有水层后加入活性炭脱色，搅拌， 过滤， 滤饼用水洗涤；
(4)、 合并所有水层后加入草酸， 搅拌、 离心， 滤饼用少量水洗涤， 再 用甲醇洗涤， 产品在 40~45°C下真空干燥， 即得拉米夫定草酸盐晶体。
上述步骤 1 中緩沖溶液为使用还原剂硼氢化钠或硼氢化钾需要的緩沖 溶液，合适的緩沖溶液例如磷酸盐或硼酸盐緩沖液，例如磷酸氢二钾水溶液， 磷酸氢二钾溶液的浓度为 20-50%,优选浓度为 25-45%,最优选的浓度为 31%。 步骤 1适宜的温度为 18 ~ 20°C。
步骤 1中适量乙醇是指原料量的 4-7倍， 优选 5-6倍。
步骤 2 中硼氢化钠 （或硼氢化钾）溶液浓度为 5mol/L, 优选采用以下 方法配制： 先配制 0.4%的氢氧化钠 （或氢氧化钾）水溶液， 然后将一定量 的硼氢化钠（或硼氢化钾）溶解在该氢氧化钠（或氢氧化钾）溶液中， 制备 浓度为 5mol/L的硼氢化钠 （或硼氢化钾）溶液。
步骤 2中 "检测至反应完全"是指经过反应后原料式 III或式 III，的量 <0.5%的投料量。
步骤 3中用于调节 pH的酸可以列举的为盐酸、 硫酸、 磷酸、 硝酸等， 优选为盐酸， 例如 6mol/L的盐酸， 调节 pH的碱可以列举的为氢氧化钠、 氢 氧化钾等。
本发明中所述的拉米夫定草酸盐均为顺式拉米夫定（拉米夫定左旋体） 与草酸形成的盐。
本发明中所述原料式 Π Γ 即 CME的化学名称为： （21，58) _5_ (4_氨基_2- 氧代 -嘧啶 _1_基) - [1， 3]氧硫杂环戊烷 _2_羧酸， 2S_异丙基 _5R -甲基 _1R -环己 酯；上述的式 I I I和 CME原料可以从市场上购买得到，或者按照 WO/1995/029174 公开的方法制备得到。
Figure imgf000007_0001
结构式： CME
本发明另一方面提供了一种以拉米夫定草酸盐为原料制备拉米夫定的方 法。
我们发现拉米夫定草酸盐容易从极性溶剂中分离出来， 从而可以用常规 的方式转变成拉米夫定游离的碱。 常规的方法包括用合适的碱， 合适的碱可 以为叔胺， 例如三乙胺。
具体为在乙醇中加入拉米夫定草酸盐， 40 ~ 45 °C下半小时内加入一定量 的三乙胺， 保温搅拌 4小时， 室温离心， 产品在 40 ~ 45 °C下真空干燥， 即得拉 米夫定晶体。
本发明还提供一种以式 111为原料制备拉米夫定的方法， 包括式 111和硼 氢化钠或硼氢化钾反应， 然后再和草酸或草酸衍生物反应， 得到拉米夫定草 酸盐， 然后通过和碱反应得到拉米夫定。
Figure imgf000008_0001
式 III 式 H 式 I
其中 R为任意取代的烷基， 包括 。烷基， 该烷基可以是直连、 支链或环 烷基， 取代基为烷基、 卤素、 羟基、 苯基等， R优选薄荷烷基、 苯基取代的薄 荷烷基， 进一步优选 L-薄荷烷基、 L-8-苯基薄荷烷基。
具体为将式 III还原并和草酸或其衍生物成盐，再游离出相应的碱得到式 I化合物， 其中式 III到式 II的步骤中反应试剂包括硼氢化钠或硼氢化钾。
本发明提供了结晶形态的拉米夫定草酸盐，提供了一种拉米夫定草酸盐 的晶体。
所述的晶体使用 Cu-Ko辐射的 X射线粉末衍射光谱、以度表示的 2 Θ基 本在， 21.04、 22.44、 23.46、 23.78、 25.26有衍射峰, 进一步在 13.94、 14.86、 21.04、 21.30、 22.44、 23.46、 23.78、 25.26、 33.36 有衍射峰， 进一步在 13.94、 14.86、 16.88、 17.66、 20.34、 21.04、 21.30、 22.44、 23.46、 23.78、 25.26、 28.30、 33.36有衍射峰。 进一步以度表示的 2 Θ在 13.94、 14.86、 16.88、 17.66、 18.74、 19.12、 19.66、 20.34、 21.04、 21.30、 22.44、 22.96、 23.46、 23.78、 25.26、 26.78、 28.30、 28.68、 28. 96、 30.08、 30.40、 31.92、 32.24、 32.84、 33.36、 34.64、 35.00、 35.80有衍射峰。
本发明的具体实施例公开了一种情形下的 X射线粉末衍射光谱，见图 1。 上述的本发明的拉米夫定草酸盐晶体是无水的、 无溶剂的， 或含^艮少或 测不出的水或溶剂， 一般本发明的晶体含小于约 1%的水或溶剂， 典型地含 有小于 0.5%的水或溶剂。本发明的晶体有可能含有小于 20%的非结晶形式， 一般本发明的拉米夫定草酸盐晶体含小于 1 0 % , 典型地含有小于 1 % , 通常 小于 0. 1 %的非结晶形态。
本发明提供的拉米夫定草酸盐晶体在约 201. 9 °C为 DSC吸热转变峰， 在约 199 °C起始吸热转变， 一个实例的 DSC图谱见图 2 , 其吸热转变温度起始温度为 199. 23 °C 0 熔点： 201. 3— 202. 7 °C。
拉米夫定的草酸盐是可药用的盐， 可以如 W091 17159所述作为抗病毒剂， 也可以配制成药物组合物。
本发明制备的拉米夫定草酸盐是一种非常出色的制备高纯度拉米夫定的 中间物质。 相比较而言， 拉米夫定的其他有机酸盐如水杨酸盐等性状发粘， 杂质含量较多， 不易干燥， 导致下一步制备高纯度的拉米夫定困难。 而拉米 夫定草酸盐为良好的晶体， 制备过程中不会出现上述现象， 通过拉米夫定草 酸盐制备的拉米夫定未经精制的情况下纯度即可达到 99. 5%以上。对于后期精 制高纯度顺式拉米夫定提供了非常有利的条件。
本发明提供的拉米夫定草酸盐为一分子草酸与两分子拉米夫定所形成的 盐， 为制备拉米夫定而脱去有机酸时所用的三乙胺量较其他拉米夫定有机酸 盐大大减少。 三乙胺是一种有毒、 易燃的化学试剂， 具有强烈的氨臭， 在空 气中微发烟， 其蒸气或液体能刺激人体皮肤和粘膜。 主要通过吸入、 食入、 经皮等方式被人体吸收， 人体不慎吸入三乙胺蒸气后， 对呼吸道有强烈的 刺激作用， 可引起肺水肿甚至死亡， 眼及皮肤接触可引起化学性灼伤。 因此减少三乙胺一方面减少了工业生产成本，更重要的是减少使用有毒试剂， 提高了生产环节的安全性和减少了最终产品的化学试剂残留。
发明人重点对比了拉米夫定草酸盐和水杨酸盐在等摩尔投料情况下脱去 有机酸所需三乙胺的用量。
1、 从拉米夫定草酸盐制备拉米夫定的方法如下:
Figure imgf000010_0001
C9H12N3〇5S
Mol. Wt.: 274.27 CsH NsOsS
Mol. Wt.: 229.26
1000ml 乙醇，加入 120克拉米夫定草酸盐（采用本发明方法制备）， 40 ~ 45 °C下在半小时内加入 60ml三乙胺， 保温搅拌 4小时， 室温离心， 产品在 40~45°C下真空干燥， 即得拉米夫定晶体。
按照公知方案采用液相色谱法对上述方法得到的拉米夫定晶体进行检 测， 仪器和色谱条件如下：
仪器： WATERS E2695四元泵； 检测器： WATERS 2489紫外吸收检测器； 工作站： Empower2软件；色谱柱： Thermo公司 BDS HYPERSILC18( 250 4.6mm); 流动相：甲醇- 0.025mol/L醋酸铵溶液（用醋酸调节 pH至 pH 3.9 ) ( 5: 95 ); 流速： 1. Oml/min; 检测波长： 277nm; 柱温： 30°C。 归一化法计算得拉米夫 定纯度达到 98 %以上， 典型为 99.6%。
2、 拉米夫定水杨酸盐制备拉米夫定的过程参见中国专利 ZL95193466.X 实施例 1 (d)部分公开的内容。
该专利说明书中记载了 66.7g拉米夫定水杨酸盐制备拉米夫定需要 26ml 三乙胺， 折算成摩尔和体积比约为 lmol： 150ml。 对比结果见表 1。 表 1: 拉米夫定草酸盐和拉米夫定水杨酸盐所需三乙胺的量之比。
Figure imgf000011_0001
从表 1可以看出以拉米夫定草酸盐为中间体生产 lmol拉米夫定可以少 用 15ml 三乙胺， 这一减少量相对于工业生产来说将大大降低三乙胺的投料 量。 如此优点一方面减少了有毒化学试剂使用带来的诸多风险， 另一方面降 低了工业生产中的原料成本。 附图说明
图 1是拉米夫定草酸盐晶体的 X-射线衍射图；
图 2是拉米夫定草酸盐晶体差示扫描量热测定图 （ DSC图） ； 具体实施方式
下面用具体实施例来进一步说明本发明的内容， 但并不以任何方式意味 着对本发明进行限制。 实施例 1
426 毫升水中加入 191.8 克磷酸氢二钾， 搅拌至完全溶解， 控温至 18 °C。加入 1L乙醇，搅拌 10分钟，再加入 140克拉米夫定原料 CME,保温 18 ~ 20°C, 在 1 小时内滴加入硼氢化钠溶液（硼氢化钠溶液配制方法： 先配制 0.4%的氢氧化钠水溶液， 然后将 28克硼氢化钠溶解在 15QmL该氢氧化钠溶 液中）， 检测至反应完全。 静置分层， 下层水层用 160mL乙醇洗涤，有机相合并后用 6mol/L盐酸调 pH至 6~6.5, 温度保持在 28~ 32°C。 再用氢氧化钠水溶液（ 1.12公斤氢氧 化钠 /14升水）调 pH至 8.0 ~ 8.3。 将乙醇蒸掉， 降至室温， 加入 280ml水溶 解用 240ml的甲苯洗涤两次， 甲苯层合并后用 70ml水萃取，合并所有水层后 加入 6克活性炭， 室温搅拌 30分钟， 过滤， 滤饼用 180ml水洗涤。 合并所有 水层后加入 25g草酸， 搅拌， 离心， 滤饼用少量水洗涤， 再用 68ml冷的甲醇 洗涤， 产品在 40~45°C下真空干燥。 即得拉米夫定草酸盐。
( 1 )核磁实验
仪器名称： Bruker AV4QQ核磁共振波谱仪; 溶剂： D20 (重水）; 1誦 R (D20) 5„: 3.18 (dd, 1H)； 3.49 (dd, 1H)； 3.79 (dd, 1H)； 3.94 (dd, 1H)； 5.25 (t 1H)； 6.03 (d, 1H); 6.24 (dd, 1H); 8.12 (d, 1H)
( 2) X射线衍射实验
仪器名称： RigakuD/Max_RCX射线衍射仪； 测试条件： 40kv 100mA; DS=SS=1 ° RS=0.3mm; 靶型： Cu; Range: 3.00-40.00; scan Rate: 10.00 Deg/min;
Figure imgf000012_0001
Figure imgf000013_0001
( 3)拉米夫定草酸盐晶型的 DSC, 其吸热转变温度起始温度为 199.23°C, 峰 值为 201.87°C,升温速率为 10.00°C/min。熔点： 201.3—202.7°C。仪器型号: waters2414。 实施例 2
1000ml乙醇中加入 120克拉米夫定草酸盐（采用本发明方法制备）， 40 ~ 45 °C下在半小时内加入 60ml三乙胺， 保温搅拌 4小时， 室温离心， 产品在 40~45°C下真空干燥， 即得拉米夫定晶体。
取 65克上步产品， 加入 1L乙醇， 升温回流 30分钟， 固体完全溶解， 加入 4 克活性炭， 继续回流 30分钟， 趁热过滤； 3到 4小时内降至室温， 保温搅拌 6 小时， 过滤， 滤饼用少量乙醇洗涤后在 40~45°C下真空干燥， 得到高纯度拉 米夫定。 纯度为 99.6%。
经测定拉米夫定熔点为： 174— 177°C。

Claims

权利要求书
1、 一种拉米夫定草酸盐， 特征在于具有如下结构式:
Figure imgf000015_0001
2、 一种结晶形态的拉米夫定草酸盐。
3、 权利要求 2所述的结晶形态的拉米夫定草酸盐， 特征在于使用 Cu-Ko辐 射， X-射线衍射以度表示的 2Θ在 21.04、 22.44、 23.46、 23.78、 25.26有 吸收峰。
4、权利要求 3所述的晶体，特征在于进一步以度表示的 2Θ在 13.94、 14.86、 21.04、 21.30、 22.44、 23.46、 23.78、 25.26、 33.36有吸收峰。
5、 权利要求 3、 4所述的拉米夫定草酸盐的晶体， 特征在于 DSC吸热转变峰 约 201.9°C, 起始转变温度为约 199°C。
6、 权利要求 1 -5任一所述的拉米夫定草酸盐的制备方法， 特征在于包括如 下步骤：
H NH2 oi^cu 丫丫
ji 0 丫丫 " /0、.、、、Ν^ #1/2?00H
RO'
S」 HO s 式 III
其中 R为任意取代的烷基， 包括 。烷基， 该烷基可以是直连、 支链或环烷 基， 取代基为烷基、 卤素、 羟基、 苯基等， 上述的草酸包括草酸衍生物， 其 中的反应试剂包括硼氢化钠或硼氢化钾， R优选薄荷烷基、 苯基取代的薄荷 烷基， 进一步优选 L-薄荷烷基、 L-8-苯基薄荷烷基。
7、 一种拉米夫定的制备方法， 该方法包括将权利要求 1 - 5中任一的拉米夫定 盐和合适的碱反应， 碱优选叔胺， 进一步优选三乙胺。
8、 一种拉米夫定的制备方法， 该方法包括如下步骤：
Figure imgf000016_0001
式 III 式 H 式 I
其中 R为任意取代的烷基， 包括 。烷基， 该烷基可以是直连、 支链或环 烷基， 取代基为烷基、 卤素、 羟基、 苯基等， R优选薄荷烷基、 苯基取代的薄 荷烷基， 进一步优选 L-薄荷烷基、 L-8-苯基薄荷烷基， 具体为将式 I I I还原并 和草酸及其衍生物成盐， 再游离出相应的碱得到式 I化合物， 其中式 I I I到式 1 1的步骤中反应试剂包括硼氢化钠或硼氢化钾。
9、 权利要求 1 - 5任一所述的拉米夫定草酸盐在制备拉米夫定中的用途。
10、 权利要求 1 - 5任一所述的拉米夫定草酸盐作为抗病毒剂的用途。
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CN102167696A (zh) 2011-08-31
KR20130021476A (ko) 2013-03-05
EP2540717B1 (en) 2015-05-20
JP5613780B2 (ja) 2014-10-29
JP2013520449A (ja) 2013-06-06
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CN102167696B (zh) 2013-09-18
US20130053560A1 (en) 2013-02-28

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