阅读说明：本技术 一种桉叶烷型倍半萜并香叶基尿黑酸甲酯杂合物及其制备方法与应用 (eudesmane type sesquiterpene geranylhomogentisate heterocomplex and preparation method and application thereof ) 是由 文松松 徐玉文 牛冲 张雷 刘琦 周广涛 窦艳丽 陈真 王雪 李琳 于 2019-10-28 设计创作，主要内容包括：本发明涉及医药技术领域,公开了一种桉叶烷型倍半萜并香叶基尿黑酸甲酯杂合物,及其制备方法与应用。该化合物分离自樟科山胡椒属植物乌药(<I>Linderaaggregata</I>)的干燥块根,命名为Linderin B。该化合物的制备方法采用了天然产物化学领域较为成熟分离纯化手段,在实际操作中容易实现。经体外生物活性研究表明Linderin B能够显著延长血浆的活化部分凝血活酶时间(activated partial thromboplastin time,APTT),可以作为抗凝血药物的先导化合物。(The invention relates to the technical field of medicines, and discloses eudesmane type sesquiterpene geranylurea-nigrosine heterozygotes, and a preparation method and application thereof Linderaaggregata ) The dried root tuber of (1) is named Linderin B. The preparation method of the compound adopts a mature separation and purification means in the field of natural product chemistry, and is easy to realize in actual operation. In vitro biological activity research shows that Linderin B can remarkably prolong the Activated Partial Thromboplastin Time (APTT) of plasma, and can be used as a lead compound of an anticoagulant.)

1. Eucalyptus alkane sesquiterpene geranylhomogentisate methyl ester heterozygote, named Linderin B, is characterized in that the chemical structural formula is as follows:

the method of making hybrids of claim 1 comprising the steps of:

(1) pulverizing dried radix Linderae root tuber to obtain coarse powder with particle size of 10 mesh or less, extracting radix Linderae root tuber coarse powder with 80% ethanol under heating and refluxing for 3 times, wherein the extraction time of times is 2 hr, and the extraction time of the second and third times is 1 hr, filtering, mixing extractive solutions, concentrating under reduced pressure, and recovering ethanol to obtain total extract;

(2) dispersing the total extract obtained in the step (1) in water, and then extracting with petroleum ether to obtain a petroleum ether phase extract;

(3) subjecting the extract obtained in the step (2) to silica gel column chromatography, petroleum ether-acetone gradient elution and thin-layer chromatography inspection to obtain seven components A-G, subjecting the component E to reversed-phase C-18 column chromatography methanol-water gradient elution, and then respectively subjecting the component E to reversed-phase C-8 acetonitrile-water isocratic elution and reversed-phase C-18 methanol-water isocratic elution to obtain eudesmane sesquiterpene geranylhomogentisate methyl ester heterozygotes (Linderin B);

the volume ratio of acetonitrile to water for the acetonitrile-water isocratic elution is 65: 35; methanol-water isocratic elution with methanol and water at a volume ratio of 70: 30.

3. The method according to claim 2, wherein the petroleum ether-acetone gradient elution of step (3) is carried out in a volume ratio of petroleum ether to acetone of 100:0 to 0: 100.

4. The method according to claim 2, wherein the methanol-water gradient elution in the step (3) is performed in a methanol-water ratio of 30:70 to 100:0 by volume.

Use of hybrids of claim 1 in the preparation of an anticoagulant.

Technical Field

The invention relates to the technical field of medicines, in particular to a traditional Chinese medicine lindera strychnifolia (lindera aggregata (Maxim.) harms of lindera of LauraceaeLindera aggregata) The preparation method and application of eudesmane sesquiterpene geranylhomogentisate heterozygotes obtained by separation.

Background

Activated Partial Thromboplastin Time (APTT) is screening tests for checking endogenous blood coagulation factors, is mainly used for checking whether deficiency of endogenous pathway blood coagulation factors VII, IX and XI exists and special inhibitors of certain factors, is a first-choice index for heparin treatment monitoring, is also a main means for blood coagulation factor treatment and lupus anticoagulant detection, and the shortening of APPT is commonly seen in thrombocytosis, prothrombotic state and thrombotic diseases, myocardial infarction, unstable angina, cerebrovascular diseases and deep vein thrombosis.

Linderae radix (A. ex Fr.) RaddeLindera aggregata) The medicine belongs to the zanthoxylum genus of Lauraceae, is a unique variety of combined spicebush root recorded in the Chinese pharmacopoeia of 2015 edition, has a long use history, and has effects of pungent and warm entering lung, spleen, kidney and bladder channels, has effects of promoting qi circulation, relieving pain, warming kidney and dispelling cold, and is used for treating diseases such as cold coagulation and qi stagnation, chest and abdomen distending pain, adverse qi, dyspnea with urgency, bladder deficiency cold, enuresis, frequent micturition, hernia pain, cold channel and abdominal pain.

Disclosure of Invention

The invention aims to provide eudesmol sesquiterpene geranylhomogentisate heterocomplexes, a preparation method thereof and application thereof in treating hemagglutination.

sesquiterpene geranylhomogentisate heterozygotes, named Linderin B, have the chemical structural formula shown below:

。

the invention also provides a preparation method of the hybrid, which comprises the following steps:

(1) pulverizing dried radix Linderae root tuber to obtain coarse powder with particle size of 10 mesh or less, extracting radix Linderae root tuber coarse powder with 80% ethanol under heating and refluxing for 3 times, wherein the extraction time of times is 2 hr, and the extraction time of the second and third times is 1 hr, filtering, mixing extractive solutions, concentrating under reduced pressure, and recovering ethanol to obtain total extract;

(2) dispersing the total extract obtained in the step (1) in water, and then extracting with petroleum ether with the same volume to obtain a petroleum ether phase extract;

(3) and (3) subjecting the extract obtained in the step (2) to silica gel column chromatography, petroleum ether-acetone gradient elution and thin-layer chromatography inspection to obtain seven components A-G, subjecting the component E to reversed-phase C-18 column chromatography methanol-water gradient elution, and then respectively subjecting the component E to reversed-phase C-8 acetonitrile-water isocratic elution and reversed-phase C-18 methanol-water isocratic elution to obtain 1 sesquiterpene-geranylhomogentisate methyl ester heterozygote, namely Linderin B.

In the petroleum ether-acetone gradient elution process in the step (3), the volume ratio of the petroleum ether to the acetone is 100:0 to 0: 100.

In the methanol-water gradient elution of the step (3), the volume ratio of methanol to water is 30:70 to 100: 0.

The volume ratio of acetonitrile to water for the acetonitrile-water isocratic elution in the step (3) is 65: 35; methanol-water isocratic elution with methanol and water at a volume ratio of 70: 30.

The invention also provides the application of the hybrid in the preparation of anticoagulant drugs.

The invention has the beneficial effects that:

1. novel compounds

The invention separates 1 cineole type sesquiterpene geranylurea-nigroate heterocomplex with novel structure from the dry root tuber of lindera strychnifolia belonging to Lauraceae, the compound is extremely rare in the natural world, the invention enriches the chemical diversity of the compound, adds 1 new compound for the compound field, and provides new evidence for the chemical taxonomic study of lindera strychnifolia.

2. The extraction method is simple

The preparation method of the compound adopts a mature technical means in the field of natural product chemistry, and is easy to realize in actual operation.

3. Treating blood coagulation

In-vitro biological activity experiments show that Linderin B can remarkably prolong the Activated Partial Thromboplastin Time (APTT) of sheep plasma and can be used as a lead compound of an anticoagulant.

Drawings

FIG. 1 high resolution mass spectrum of Compound Linderin B;

FIG. 2 the hydrogen spectrum of compound Linderin B;

FIG. 3 carbon and DEPT135 spectra of Compound Linderin B;

FIG. 4 of Compound Linderin B¹H-¹H COSY spectrum;

FIG. 5 HSQC spectra of compound Linderin B;

FIG. 6 HMBC spectra of compound Linderin B;

FIG. 7 NOESY spectrum of Compound Linderin B;

FIG. 8 ultraviolet absorption spectrum of Compound Linderin B;

FIG. 9 circular dichroism spectrum of Compound Linderin B;

FIG. 10 Infrared spectrum of Compound Linderin B.

Detailed Description