CN111808051A - New compound and application thereof in preparing medicine for treating bone fracture - Google Patents

Abstract

The invention discloses a novel compound and application thereof in preparing a medicament for promoting fracture healing, wherein the structural formula of the compound is as follows:

the compound has obvious effect of promoting fracture healing and can be used for preparing medicaments for treating fracture.

Description

New compound and application thereof in preparing medicine for treating bone fracture

Technical Field

The invention belongs to the technical field of medicines, and particularly relates to a novel compound and application thereof in preparing a medicine for treating fracture.

Background

With the rapid development of industry, transportation industry and building industry, accidents are more frequent. And the social aging is increasingly serious, and osteoporosis is more and more common in clinic, so that fracture patients are more and more promoted. The fracture is slow to heal, the action of the patient is limited, and the life quality of the patient is seriously influenced.

The clinical treatment scheme of the fracture is reduction, fixation and functional exercise. In the aspect of medicament treatment, the Chinese patent medicines are mainly taken to play the roles of promoting blood circulation to remove blood stasis, and relieving swelling and pain. However, the Chinese patent medicine has complex components, the effective components and the action base are not clear, and the clinical curative effect varies from person to person. The finding of monomer chemical components capable of promoting fracture healing from traditional Chinese medicines is a hot point of research in recent years, and is also a direction for developing new medicines for treating fracture.

The kudzu root is the dry root of Pueraria lobata Ohwi of Leguminosae, is a traditional Chinese medicine, and has the effects of relieving muscles and fever, promoting eruption, promoting the production of body fluid to quench thirst, invigorating yang and relieving diarrhea. The traditional Chinese medicine composition is usually used for treating symptoms such as exterior syndrome fever, strong neck and back pain, measles without adequate eruption and the like, and is often shown in some prescription groups for treating fracture, modern pharmacology indicates that the kudzuvine root has the effects of promoting bone formation and resisting osteoporosis, but the effective components are not clear.

Disclosure of Invention

Aiming at the defects of the prior art, the invention aims to separate a new compound from kudzuvine root and develop the new compound into a medicinal preparation for treating fracture.

The preparation method of the novel compound comprises the following steps:

(1) extracting radix Puerariae with 70% ethanol, and concentrating the extractive solution until no ethanol smell exists;

(2) sequentially extracting the concentrated solution with petroleum ether, dichloromethane and ethyl acetate, and concentrating the extract under reduced pressure to obtain extract, respectively petroleum ether extract, dichloromethane extract and ethyl acetate extract;

(3) performing silica gel column chromatography on the ethyl acetate extract, eluting with a mixed solution of dichloromethane and methanol in a volume ratio of 50:1, discarding the eluent, eluting with a mixed solution of dichloromethane and methanol in a volume ratio of 10:1, collecting the eluent, and concentrating to obtain an extract;

(4) dissolving the extract with methanol, and performing preparative liquid phase separation under the following chromatographic conditions: octadecylsilane chemically bonded silica is used as a filling agent, a mobile phase is methanol to water with a volume ratio of 35:65, an ultraviolet detector detects the wavelength of 320nm, chromatographic peak eluent with retention time of 135-plus of 147 minutes is collected, and a solvent is recovered, so that the target compound is obtained.

The new compound is used for preparing the medicine for treating the bone fracture.

Drawings

FIG. 1 is an X-ray film for the test of the new compound of the present invention for promoting fracture healing

FIG. 2 shows the hydrogen nuclear magnetic resonance spectrum of the novel compound of the present invention

FIG. 3 shows the NMR spectrum of the novel compound of the present invention

FIG. 4 shows the NMR HMBC spectra of the novel compounds of the present invention

FIG. 5 shows the NMR HSQC spectra of the novel compounds of the present invention

Detailed Description

The present invention is further illustrated by the following examples, which are not intended to limit the practice of the invention.

Example 1

New compound effect of promoting fracture healing

1. Materials and methods

1.1 animal model 20 healthy New Zealand white immune of 6 months, each half of male and female, body weight 2.5 + -0.5 kg, divided into 4 groups according to body weight, namely control group (simple fracture group), low dose group, middle dose group, high dose group, each group comprising 5 animals. All animals were injected intravenously with 50mg/kg of sodium pentobarbital, the left forelimb was clipped, sterilized, and the skin was surgically opened, the subcutaneous tissue was separated, and the ulna was exposed after anesthesia. Incomplete fracture of the middle ulna is caused by a steel saw. Intramuscular injection of penicillin sodium salt within three days after surgery prevented infection.

1.2 administration by intragastric administration, the new compound of the invention is added with 50% ethanol to prepare a solution of 100 mug/ml. The low dose group is administered at a dose of 100 μ g/kg^-1·d^-1The administration dosage of the medium dose group is 300 mu g kg^-1·d^-1The high dose group is administered at a dose of 900 μ g/kg^-1·d^-1The blank group was drenched with 10ml of distilled water. Animals were sacrificed with continuous dosing.

1.3 Observation index all animals were sacrificed two weeks after surgery, the forearms were taken for X-ray film, and the fracture healing condition was observed.

2. Results

The X-ray film result shows that compared with the control group, the fracture healing speed of each group of the novel compound is accelerated, the bone formation is active and is dose-dependent, the callus formation of the high-dose group is obvious, and the fracture line is fuzzy, which is shown in figure 1.

Example 2

(1) Extracting 1kg of radix Puerariae with 70% ethanol, and concentrating the extractive solution until no ethanol smell exists;

(2) sequentially extracting the concentrated solution with petroleum ether, dichloromethane and ethyl acetate, and concentrating the extract under reduced pressure to obtain extract, respectively petroleum ether extract, dichloromethane extract and ethyl acetate extract;

(3) performing silica gel column chromatography on the ethyl acetate extract, taking 500g of silica gel, performing wet column packing, performing dry sample loading, eluting by 5L of a mixed solution of dichloromethane and methanol with a volume ratio of 50:1, discarding the eluent, eluting by 10L of a mixed solution of dichloromethane and methanol with a volume ratio of 10:1, collecting the eluent, and concentrating to obtain an extract;

(4) dissolving the extract with methanol, and performing preparative liquid phase separation under the following chromatographic conditions: octadecylsilane chemically bonded silica is used as a filling agent (20X 250mm, 10 mu m), a mobile phase is methanol to water with a volume ratio of 35:65, an ultraviolet detector detects that the wavelength is 320nm, the flow rate is 10ml per minute, chromatographic peak eluent with retention time of 135-147 minutes is collected, and the solvent is recovered, so that the target compound is obtained.

Example 3

Structure confirmation of the Compound described in the present invention

White amorphous powder, dissolved in methanol. HR-ESI-MS M/z [ M + H ]]⁺413.1238 (calculated 413.1236), and recombination¹H-NMR spectrum and¹³the molecular formula of the compound is presumed to be C through C-NMR spectrum data₂₂H₂₀O₈. Binding by HSQC and HMBC Spectroscopy¹H-NMR and¹³the information given by the C-NMR spectra, which is shown in Table 1, attributes the hydrocarbon signals of the novel compounds of the present invention.¹The H-NMR spectrum is shown in figure 2,¹³the C-NMR spectrum is shown in FIG. 3, the HMBC spectrum is shown in FIG. 4, and the HSQC spectrum is shown in FIG. 5. Through [ alpha ]]_D-73.5°(acetone；c 0.50)，CD(MeOH)[θ](nm):-1.5×10⁴(320)，-2.5×10⁴(280)，+1.8×10⁴(250)，-0.6×10⁴(230) The absolute configuration of the C-4 position is determined. In summary, the chemical structure of the novel compound of the present invention is:

TABLE 1¹H(600MHz)and¹³C(150MHz)NMR Spectroscopic Data in DMSO-d₆(inppm,Jin Hz).

Claims (3)

1. A novel compound characterized by the following chemical structure:

2. a process for the preparation of the novel compounds of claim 1, characterized in that: the method comprises the following steps:

(1) extracting radix Puerariae with 70% ethanol, and concentrating the extractive solution until no ethanol smell exists;

(2) sequentially extracting the concentrated solution with petroleum ether, dichloromethane and ethyl acetate, and concentrating the extract under reduced pressure to obtain extract, respectively petroleum ether extract, dichloromethane extract and ethyl acetate extract;

(3) performing silica gel column chromatography on the ethyl acetate extract, eluting with a mixed solution of dichloromethane and methanol in a volume ratio of 50:1, discarding the eluent, eluting with a mixed solution of dichloromethane and methanol in a volume ratio of 10:1, collecting the eluent, and concentrating to obtain an extract;

(4) dissolving the extract with methanol, and performing preparative liquid phase separation under the following chromatographic conditions: octadecylsilane chemically bonded silica is used as a filling agent, a mobile phase is methanol to water with a volume ratio of 35:65, an ultraviolet detector detects the wavelength of 320nm, chromatographic peak eluent with retention time of 135-plus of 147 minutes is collected, and a solvent is recovered, so that the target compound is obtained.

3. The novel compounds according to claim 1, for the preparation of a medicament for the treatment of bone fractures.

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