CN107050428B

CN107050428B - FGF20 medicament and application thereof in treatment of cerebral trauma

Info

Publication number: CN107050428B
Application number: CN201710176647.3A
Authority: CN
Inventors: 林丽; 陈珺; 王雪; 黄文婷; 田海山; 李校堃
Original assignee: Wenzhou Medical University
Current assignee: Wenzhou Medical University
Priority date: 2017-03-23
Filing date: 2017-03-23
Publication date: 2020-05-05
Anticipated expiration: 2037-03-23
Also published as: CN107050428A

Abstract

The invention belongs to the technical field of protein/polypeptide medicines, and provides a medicinal composition for treating brain trauma, which comprises a fibroblast growth factor 20 and can be used for repairing the blood brain barrier of the injured brain. In addition, the invention also provides a pharmaceutical preparation comprising the pharmaceutical composition, application and the like.

Description

FGF20 medicament and application thereof in treatment of cerebral trauma

Technical Field

The invention belongs to the technical field of protein/polypeptide medicines, and particularly relates to a pharmaceutical composition containing FGF20, which is used for treating brain trauma, especially repairing the blood brain barrier of the injured brain. In addition, the invention also relates to a therapeutic application and the like comprising the pharmaceutical composition.

Background

Brain trauma, also known as Traumatic Brain Injury (TBI), refers to the damage of brain tissue caused by impact or impact of external mechanical force on the head, rapid acceleration or deceleration, penetration of foreign objects into the brain, etc., resulting in temporary or permanent cognitive, physical and psychological dysfunction, etc. Brain trauma is not a single pathophysiological mechanism but a complex disease process, and no effective treatment means exists at present.

The Blood brain barrier (Blood brain barrier) is composed of brain microvascular endothelial cells, astrocyte end-feet, pericytes and a basement membrane, is not a simple physical barrier, and plays an important role in maintaining the homeostasis of brain microenvironment. Among these, the first line of defense is endothelial cells, which present a dynamic, highly regulated interface between the blood and the central nervous system. The blood brain barrier endothelial cells are distinguished from peripheral endothelial cells mainly by their lack of a fenestration, minimal cellular drinking water and the presence of tight junctions between endothelial cells. The main role of the blood-brain barrier is to tightly regulate the rate of cell permeation. This is mediated primarily by tight junctions between capillary endothelial cells, restricting movement of cytosol, ions and water.

The protein medicine (also called active growth factor group and nutrient component) in the prior art not only has complex components, but also has large dosage, is not easy to produce stably, and is difficult to popularize and apply in practice. In particular, the functions and structures of different members of the existing Fibroblast Growth Factor (FGF) family are different, and at present, 23 members exist, and it can be confirmed that selection of one member is not easy.

In summary, although there is no effective treatment for brain trauma at present, the present inventors have not retracted from the difficulties of the prior art, and after long-term and arduous studies, especially in combination with the experience of the present inventors, have found that an FGF (i.e., FGF20) is effective in treating brain trauma, and especially in repairing the blood brain barrier of injured brain.

Disclosure of Invention

The invention aims to provide a pharmaceutical composition for treating brain trauma, in particular for repairing blood brain barrier of the injured brain. In addition, the invention also provides a pharmaceutical preparation comprising the pharmaceutical composition, application and the like.

In particular, in a first aspect, the present invention provides a pharmaceutical composition for the treatment of brain trauma comprising fibroblast growth factor 20(FGF 20).

FGF20 is known and the prior art is keen to study non-natural mutants of FGF20, whereas the present inventors have found that human native FGF20 (i.e. non-mutated FGF20) is more effective, and thus in a specific embodiment of the present invention FGF20 is human native FGF20, which can be prepared as described in chinese patent application CN 104293821A.

Preferably in the pharmaceutical composition of the first aspect of the invention, the pharmaceutically active ingredient comprises FGF20 only. That is, the pharmaceutical composition of the first aspect of the present invention does not comprise other pharmaceutically active ingredients for treating brain trauma, other than FGF 20.

Preferably in the pharmaceutical composition of the first aspect of the invention, the treatment of brain trauma is the repair of the blood brain barrier of a wounded brain.

Also preferably in the pharmaceutical composition of the first aspect of the invention, the wound is a hydraulic wound.

In a second aspect, the present invention provides a pharmaceutical formulation for use in the treatment of brain trauma comprising a pharmaceutical composition according to the first aspect of the invention and a pharmaceutically acceptable excipient. Pharmaceutically acceptable excipients in this context refer to nontoxic fillers, stabilizers, diluents, carriers, solvents or other formulation excipients. For example, diluents, excipients, such as microcrystalline cellulose, mannitol, and the like; fillers, such as starch, sucrose, and the like; binders, such as starch, cellulose derivatives, alginates, gelatin and/or polyvinylpyrrolidone; disintegrants, such as calcium carbonate and/or sodium bicarbonate; absorption promoters, such as quaternary ammonium compounds; surfactants such as cetyl alcohol; carriers, solvents, such as water, physiological saline, kaolin, bentonite, etc.; lubricants, such as talc, calcium/magnesium stearate, polyethylene glycol, and the like.

Preferably, in the pharmaceutical preparation of the second aspect of the present invention, the pharmaceutically acceptable excipient is an injectable pharmaceutical preparation excipient. That is, the pharmaceutical formulation of the second aspect of the present invention is an injectable (pharmaceutical) formulation. For example, the pharmaceutical composition of the first aspect of the present invention may be dissolved or diluted with physiological saline, and configured as an aqueous injection; or mixing with mannitol and human serum albumin, lyophilizing, and making into injection (lyophilized) powder.

Preferably in the pharmaceutical formulation of the second aspect of the invention, the pharmaceutically active ingredient is delivered solely by the pharmaceutical composition of the first aspect of the invention. That is, the pharmaceutical formulation of the second aspect of the present invention consists of the pharmaceutical composition of the first aspect of the present invention and pharmaceutically acceptable excipients.

The inventors have found that only one injection of an effective amount of FGF20 is required to be effective, and therefore it is preferred that the pharmaceutical formulation of the second aspect of the invention comprises an effective amount of FGF 20. The present inventors found that the dose of the drug is 0.6 to 2.4mg/kg, preferably 1.2 to 1.8mg/kg, and most preferably 1.5mg/kg in rats, and the effective amount of the drug can be converted into rats according to the body weight of the rats. In this context, an effective amount refers to an effective amount for an adult, depending on the context, if not otherwise specified. The dose of an adult can be derived from the dose of an experimental animal according to the equivalent dose conversion relationship between the experimental animal and a human (generally, refer to the guidance of drug administration such as FDA and SFDA, and refer to Huang Tanhua, et al. equivalent dose conversion between animals and a human body in pharmacological experiments, Chinese clinical pharmacology and therapeutics, 2004, 9 (9): 1069-1072) known by those skilled in the art. Specifically, in the case of rats, which are experimental animals used in the embodiment of the present invention, the conversion relationship with adults is generally 6: 1, the calculated dose for an adult is 0.1 to 0.4mg/kg, preferably 0.2 to 0.3mg/kg, and most preferably 0.25 mg/kg. From the normal adult body weight of 60Kg, it can be deduced that the pharmaceutical preparation of the second aspect of the present invention preferably comprises 6-24 mg of FGF20, preferably 12-18 mg of FGF20, and most preferably 15mg of FGF 20.

Preferably in the pharmaceutical formulation of the second aspect of the invention, the treatment of brain trauma is the repair of the blood-brain barrier of a wounded brain.

Also preferably in the pharmaceutical formulation of the second aspect of the invention, the wound is a hydraulic wound.

In a third aspect, the present invention provides the use of FGF20 in the manufacture of a medicament for the treatment of brain trauma.

Preferably, in the use according to the third aspect of the invention, FGF20 is used alone as the pharmaceutically active ingredient. That is, the third aspect of the present invention preferably provides the use of FGF20 alone as a pharmaceutically active ingredient for the preparation of a medicament for the treatment of brain trauma.

Preferably, in the third aspect of the present invention, the medicament comprises 6-24 mg of FGF20, preferably 12-18 mg of FGF20, and most preferably 15mg of FGF 20.

Preferably in the use of the third aspect of the invention, the treatment of brain trauma is the repair of the blood brain barrier of a wounded brain.

Also preferably in the use of the third aspect of the invention, the wound is a hydraulic wound.

Preferably in the use of the third aspect of the invention, the medicament is an injectable medicament, such as a pharmaceutical formulation according to the second aspect of the invention.

The invention has the following beneficial effects: the medicine can be used for treating brain trauma, in particular repairing blood brain barrier of the injured brain; the medicine only needs conventional injection, does not need brain injection, can take effect only by one-time injection, and is beneficial to actual popularization and application; the medicine has remarkable therapeutic effect, and can especially improve behavioral function reduction caused by brain trauma.

The invention will be described in detail below by means of specific embodiments and the accompanying drawings. It is to be expressly understood that the description is illustrative only and is not intended as a definition of the limits of the invention. Many variations and modifications of the present invention will be apparent to those skilled in the art in light of the teachings of this specification.

Drawings

FIG. 1 shows a photograph of staining for changes in blood brain barrier permeability as detected by Evans Blue (Evans Blue) staining.

Fig. 2 shows the quantitative results of the blood brain barrier permeability change detected by evans blue staining, wherein the ordinate represents the amount of evans blue extracted per mg of brain tissue (ng), wherein the differential significance symbols: p <0.01, compared to Sham group (Sham); # P <0.05, compared to model group (TBI).

Fig. 3 shows the result of the Cylinder behavior scoring, where the symbols of significance of difference: p <0.01, compared to Sham group (Sham); # P <0.05, compared to model group (TBI).

Detailed Description

Hereinafter, specific embodiments of the present invention will be described in detail. It should be noted that technical features or combinations of technical features described in the following embodiments should not be considered as being isolated, and they may be combined with each other to achieve better technical effects.

Example 1 establishment of Hydraulic brain trauma rat model

Example 2 treatment of Hydraulic brain trauma rats with FGF20

Rats were randomized into three groups: 1, Sham group (Sham): the rest treatment is the same as the model group except that the hydraulic impact damage is not carried out; model group (TBI): a rat right side brain trauma model was established as in example 1; 3, FGF20 administration group (TBI + FGF 20): after 0.5h of establishing the right side brain trauma model of rats as in example 1, FGF20 was injected i.p. at a rat dose of 1.5 mg/kg.

After the rats stand on the right side brain trauma model for 24h, evans blue (4%, 3mL/kg) is injected through the tail vein, and the whole brain is taken 3h after injection, and the staining condition of evans blue in the injured area is photographed (figure 1). The brain was then soaked in an equal volume of trichloroacetyl overnight and the supernatant centrifuged at 620nm to determine the evans blue content (fig. 2). The results are shown in fig. 1 and 2, indicating that FGF20 can significantly reduce the amount of evans blue that passes the blood-brain barrier, indicating that FGF20 has a repairing effect on the integrity of the blood-brain barrier.

The remaining rats were subjected to the Cylinder neural score and the brain injury in the rats was examined behaviorally (fig. 3). The behavioral score shows that the model group scores 32 +/-2.2, and the FGF20 administration group scores 18.7 +/-2.1, and the FGF20 can remarkably improve the behavioral function reduction caused by the hydraulic brain trauma.

Claims

Use of FGF20 alone as pharmaceutically active ingredient for the preparation of a medicament for repairing the blood-brain barrier of a wounded brain, wherein the pharmaceutically active ingredient of said medicament comprises FGF20 alone.
2. The use of claim 1, wherein the wound is a hydraulic wound.
3. The use of claim 1, wherein the medicament is an injectable medicament.