CN112121050B

CN112121050B - Application of ATF4 inhibitor in preparation of drug for preventing or treating ankylosing spondylitis

Info

Publication number: CN112121050B
Application number: CN202010962559.8A
Authority: CN
Inventors: 沈慧勇; 马梦君; 杨文�; 李鸿宇; 吴燕峰; 王鹏; 米汝佳; 卢艺萱
Original assignee: Eighth Affiliated Hospital of Sun Yat Sen University
Current assignee: Eighth Affiliated Hospital of Sun Yat Sen University
Priority date: 2020-09-14
Filing date: 2020-09-14
Publication date: 2022-07-29
Anticipated expiration: 2040-09-14
Also published as: CN112121050A

Abstract

The invention relates to the field of medical biology, in particular to application of an ATF4 inhibitor in preparation of a medicine for preventing or treating ankylosing spondylitis. At present, AS treatment cannot solve the problem of pathological osteogenic fusion of joints and spines of patients, and in order to prevent the occurrence of joint rigidity and spine deformity, the invention finds a new AS treatment target; compared with the existing AS treatment technology, the invention can effectively inhibit AS focus angiogenesis and pathological osteogenesis, and break through the bottleneck of AS treatment.

Description

Application of ATF4 inhibitor in preparation of drug for preventing or treating ankylosing spondylitis

Technical Field

The invention relates to the field of medical biology, in particular to application of an ATF4 inhibitor in preparation of a medicine for preventing or treating ankylosing spondylitis.

Background

Ankylosing Spondylitis (AS) is a disease with inflammation of the sacroiliac joint and spinal attachment points AS the main symptoms. At present, the etiology is not clear, and no root treatment method exists. Treatment of AS is primarily to control inflammation, alleviate or relieve symptoms, maintain normal posture and optimal functional position, and prevent deformity. The treatment is divided into drug treatment, non-drug treatment and operation treatment. Non-drug therapy includes maintaining normal posture, such as sitting and standing with chest support and abdomen support; proper amount of physical exercise; keeping optimistic mood, stopping smoking, etc. The drug therapy is preferably non-steroidal anti-inflammatory drugs (NSAIDs), and other drug therapies include tumor necrosis factor (TNF-alpha) antagonists and disease-modifying antirheumatic drugs (DMARDs), among others. Serious kyphosis and deformity of spine, and operation correction is performed after the disease state is stable, and the kyphosis person can correct kyphosis by means of spine osteotomy.

Ankylosing spondylitis is difficult to diagnose early, causes of diseases are not clear up to now, the existing treatment method can only relieve inflammation, and along with the progress of diseases, patients can suffer from tendon onset and termination inflammatory pain and low back pain; the movement of joints and spines is limited, and the serious patient has joint rigidity and pathological spine osteogenesis fusion deformity.

Disclosure of Invention

The invention aims to overcome the defects of the prior art, provides the application of an ATF4 inhibitor in preparing a medicament for preventing or treating ankylosing spondylitis, and finds a new AS treatment target point for preventing ankylosis and spinal deformity.

In order to achieve the purpose, the invention adopts the technical scheme that: provides an application of an ATF4 inhibitor in preparing a medicament for preventing or treating ankylosing spondylitis.

Through preliminary studies, ATF4 in Mesenchymal Stem Cells (MSC) of AS patients regulates and controls SMURF2 to be up-regulated, and regulates and controls lesion synovial angiogenesis abnormality, so that pathological osteogenesis is caused. By injecting ATF4 inhibitor into tail vein of AS mouse model, angiogenesis at joint synovium part of mouse is reduced obviously, and pathological bone formation of spine is controlled effectively. The ATF4 inhibitor is suggested to be a new therapeutic target of AS.

As a preferred embodiment of the use of the invention, the ATF4 inhibitor is KIRA-7, and the KIRA-7 has the molecular formula C ₂₇ H ₂₃ FN ₆ O。

As a preferred embodiment of the use according to the invention, the medicament is for subcutaneous, intravenous, intramuscular or nasal administration.

The invention also provides a pharmaceutical composition for treating ankylosing spondylitis, which comprises the ATF4 inhibitor.

As a preferred embodiment of the pharmaceutical composition of the present invention, the ATF4 inhibitor is KIRA-7, and the molecular formula of the KIRA-7 is C ₂₇ H ₂₃ FN ₆ O。

As a preferred embodiment of the pharmaceutical composition of the present invention, the pharmaceutical composition further comprises a pharmaceutically acceptable carrier.

The invention has the beneficial effects that:

at present, AS treatment cannot solve the problem of pathological osteogenic fusion of joints and spines of patients, and in order to prevent the occurrence of joint rigidity and spine deformity, the invention finds a new AS treatment target; compared with the existing AS treatment technology, the invention can effectively inhibit AS focus angiogenesis and pathological osteogenesis, and break through the bottleneck of AS treatment.

Drawings

FIG. 1: the invention relates to an experimental flow chart.

FIG. 2: immunofluorescence CD31, OSX staining contrast.

FIG. 3: immunohistochemical OSX staining contrast.

FIG. 4: micro-CT shows a contrast map of the generation of the ankle joint calcaneus osteophyte of the mouse.

Detailed Description

To more clearly illustrate the technical solutions of the present invention, the following embodiments are further described, but the present invention is not limited thereto, and these embodiments are only some examples of the present invention.

Example 1

(1) AS mouse animal model construction

Female BALB/c mice of 24 weeks old are selected and are injected with Proteoglycan (Proteoglycan) in the abdominal cavity (1 time for each week of 0, 3 and 6, 3 times for each injection, and 100ug for each injection) to obtain the AS mouse animal model.

(2) Group administration

The AS mice obtained in step (1) were randomly divided into three groups including a control group, a TNF-a antagonist group and an ATF4 inhibitor (KIRA-7, imidazopyrazine compound) group, and the three groups were administered physiological saline, a TNF-a antagonist and an ATF4 inhibitor (KIRA-7, C, 4) respectively, from week 10 ₂₇ H ₂₃ FN ₆ O, imidazopyrazine compounds) 5mg/kg each was injected in tail vein once every 4 weeks for 6 total injections by week 30.

(3) Detecting angiogenesis and pathological osteogenesis conditions of mice

Mice in three groups were sacrificed using overdose anesthesia, starting at week 14, once every 4 weeks for a total of 6 times at week 34.

Taking 3 rd to 5 th lumbar vertebra segments of a mouse spine, sequentially fixing, decalcifying, embedding in paraffin, slicing, HE staining and IF staining (CD31 and OSX) to detect the pathological osteogenesis condition of the mouse spine.

As shown in FIG. 2, CD31 is a marker for angiogenesis and OSX is a marker for bone formation. Immunofluorescence of spinal sections of mice in the ATF4 inhibitor group shows that CD31 and OSX staining are obviously weakened compared with those of a normal saline group and a TNF-alpha inhibitor group, and the result shows that the ATF4 inhibitor can obviously inhibit angiogenesis and pathological osteogenesis of AS mice.

And secondly, taking the 3 rd to 5 th lumbar vertebra segment of the mouse spine, sequentially fixing, decalcifying, embedding in paraffin, slicing, and detecting pathological osteogenesis of the mouse spine by IHC (OSX).

The results are shown in fig. 3, and OSX staining of spinal sections of mice in the ATF4 inhibitor group is obviously weakened compared with that of the normal saline group and the TNF-alpha inhibitor group, which indicates that the ATF4 inhibitor can obviously inhibit pathological osteogenesis of AS mice.

And thirdly, after the mice are killed, micro-CT scanning is carried out on the ankle joints of the mice, and the generation condition of the calcaneal osteophyte is detected and evaluated.

The results are shown in fig. 4, where miicro-CT showed significantly less heel osteophyte formation in the ATF4 inhibitor group than in the normal saline group and the TNF- α inhibitor group, indicating that ATF4 inhibitor can inhibit AS mouse osteophyte formation.

Fourthly, grading pathological osteogenesis of the mice in each group, wherein the grading standard of the pathological osteogenesis is as follows: 0 minute: normal; 1 minute: slight hyperplasia of vertebral body edge; and 2, dividing: obvious osteophyte formation at the edge of the vertebral body; and 3, dividing: a bone bridge is formed at the edge of one side of the vertebral body; and 4, dividing: the edges of the vertebral bodies at both sides form a bone bridge and interbody fusion.

The results are shown in Table 1, the pathological spinal bone formation score was significantly reduced at week 34 of the KIRA-7 experiment compared with week 14, which was only 16.7% of week 14, while the pathological spinal bone formation scores of the control group and the TNF-alpha group were not significantly improved, indicating that KIRA-7 can relieve the pathological bone formation symptoms of AS mice.

TABLE 1 spinal pathological osteogenesis score

	Control group	TNF-alpha group	KIRA-7 group
				Week 14	4.2±0.6	4.1±0.9	4.2±1.6
Week 18	4.6±0.7	4.3±1.1	3.5±0.8
				At 22 weeks	5.0±1.2	4.7±1.8	2.7±1.2
At 26 th week	5.7±0.9	5.3±0.6	2.0±0.6
				At week 30	7.2±1.6	5.7±1.1	1.3±0.8
Week 34	7.9±2.0	6.1±1.4	0.7±0.3

In conclusion, compared with the physiological saline group and the TNF-alpha antagonist group, the ATF4 inhibitor group has stronger focal angiogenesis inhibiting and pathological bone formation inhibiting capabilities than the physiological saline group and the TNF-alpha antagonist group; the TNF-alpha antagonist group has slightly stronger capability of inhibiting the lesion angiogenesis and pathological osteogenesis of AS mice than the normal saline group. No side effects such as abnormal death or infection of the three groups of mice are found.

Finally, it should be noted that the above embodiments are only used for illustrating the technical solutions of the present invention and not for limiting the protection scope of the present invention, and although the present invention is described in detail with reference to the preferred embodiments, it should be understood by those skilled in the art that modifications or equivalent substitutions can be made on the technical solutions of the present invention without departing from the spirit and scope of the technical solutions of the present invention.

Claims

The application of an ATF4 inhibitor in preparing a medicament for preventing or treating ankylosing spondylitis; the ATF4 inhibitor is KIRA-7, and the molecular formula of the KIRA-7 is C ₂₇ H ₂₃ FN ₆ O。
2. The use according to claim 1, wherein the medicament is for subcutaneous, intravenous, intramuscular or nasal administration.