WO2019148471A1 - Novel use, medicament and health supplement using fg-4592 or salt thereof - Google Patents

Abstract

The present invention relates to a novel use, medicament, and health supplement using FG-4592. The novel use is the use of FG-4592 or a pharmaceutically acceptable salt thereof in the preparation of a product for treating and/or preventing neurological impairment diseases or neurodegenerative diseases. The product can be a medicament and/or health supplement. The invention overcomes the limitation that current FG-4592 is only used for the treatment of anemia. The FG-4592 was able to significantly increase the spatial memory function of APP/PS1 transgenic mice, reducing the decline of cognitive function, and can be used for the treatment of early-stage cognitive impairment, stroke, Alzheimer's disease, cognitive dysfunction, vascular dementia, Parkinson's disease, etc.

Description

New uses of FG-4592 or its salts, as well as pharmaceuticals and health products Technical field

The present invention relates to the field of medicine and health care products, in particular to new uses of FG-4592, as well as pharmaceuticals and health care products, in particular to FG-4592 or a pharmaceutically acceptable salt thereof for the preparation of a therapeutic and/or prophylactic neurological disease or neurodegenerative Use in sexually transmitted diseases products.

Background technique

Cognition is the process by which the brain accepts external environmental information, processes it, transforms it into an intrinsic psychological activity, and acquires knowledge or application skills, including memory, language, visual space, execution, calculation, and understanding. Cognitive impairment refers to the impairment of one or more of the above cognitive functions and affects the individual's daily life and social skills. Alzheimer's disease affects more than 1.5 million people worldwide and is the most common cause of cognitive dysfunction. However, there are currently no clear and effective prevention and treatment measures for cognitive dysfunction caused by Alzheimer's disease.

Roxadustat, also known as FG-4592, has the molecular formula C19H16N205, the chemical name is N-[(4-hydroxy-1-methyl-7-phenoxy-3-isoquinolinyl)carbonyl]glycine, CAS No. 808118-40- 3, its structural formula is as follows:

FG-4592由美国FibroGen公司原研，目前与阿斯利康公司合作开发的口服缺氧诱导因子(HIF)脯氨酰羟化酶(PHDs)抑制剂，本品可用于治疗慢性肾损伤等引起的贫血，目前在美国 FDA处于3期临床研究阶段。

FG-4592 is an anti-hypoxia-inducible factor (HIF) prolyl hydroxylase (PHDs) inhibitor developed by FibroGen, USA, and is currently being developed in collaboration with AstraZeneca. This product can be used to treat anemia caused by chronic kidney injury. Currently, the US FDA is in Phase 3 clinical research.

FG-4592 is a proline hydroxylase (PHDs) inhibitor that reduces the degradation of hypoxia-inducible factor 1α (HIF-1α) by inhibiting proline hydroxylase, thereby activating HIF signaling pathway and promoting downstream of HIF. Expression of functional genes. Hypoxia-inducible factors can regulate the expression of a variety of hypoxia-related genes and participate in the regulation of angiogenesis, erythropoiesis, glucose metabolism, cell survival, and inflammatory responses. FG-4592 stabilizes HIF-1α by inhibiting proline hydroxylase activity, activates HIF signaling, and causes anemia in a variety of factors (Becker, 2017), spinal cord injury (Wu, 2016), liver injury (Meent, 2016) ), radiation damage (Colbert, 2016) plays a protective role.

The invention patent CN106187888 discloses FG-4592 single crystal and a preparation method thereof, and the patent specification states that FG-4592 is an oral hypoxia-inducible factor (HIF) prolyl hydroxylase inhibitor, which is clinically used for treatment. Anemia, FG-4592 is the most distant candidate for clinical development of potential anemia treatments. This patent document also only discloses the use of FG-4592 for the treatment of anemia.

However, whether FG-4592 has been able to activate HIF signaling pathway and improve cognitive dysfunction has not been reported so far.

Summary of the invention

The present invention provides the use of FG-4592, or a pharmaceutically acceptable salt thereof, for the manufacture of a medicament for the treatment and/or prevention of a neurological or neurodegenerative disease.

The nerve injury disease is early cognitive impairment and stroke.

The neurodegenerative disease is one or more of Alzheimer's disease, cognitive dysfunction, vascular dementia, and Parkinson's disease.

The product is a pharmaceutical and/or health supplement.

The product is capable of enhancing the spatial memory function of a human or an animal.

The present invention also provides a pharmaceutical for treating and/or preventing the above-mentioned nerve damage disease or neurodegenerative disease, wherein the active ingredient is FG-4592 or a pharmaceutically acceptable salt thereof.

The drug also contains one or more pharmaceutically acceptable excipients or carriers.

The adjuvant or carrier includes conventional diluents, excipients, fillers, binders, wetting agents, disintegrating agents, absorption enhancers, surfactants, adsorption carriers, lubricants in the pharmaceutical field.

The pharmaceutical preparation can be prepared from a tablet, a capsule, an effervescent tablet, a granule, a powder, a dispersible tablet, an oral solution, a pill or an injection.

The present invention also provides a health-care product for treating and/or preventing the above-mentioned nerve damage disease or neurodegenerative disease, wherein the active ingredient is FG-4592 or a pharmaceutically acceptable salt thereof.

The health care product also contains one or more excipients or carriers that are adapted to different dosage forms.

The health care product can be formulated into a tablet, a capsule or an oral solution.

The above-mentioned medicine or health care product provided by the present invention may only use FG-4592 and a pharmaceutically acceptable salt thereof as a medicinal ingredient or an active ingredient, or may be combined with two or more other compounds or pharmaceutically acceptable salts thereof. Active ingredient or active ingredient.

FG-4592 can significantly increase APP (β amyloid precursor protein) / PS1 (presenilin 1) transgenic mice under intraperitoneal administration (APP/PS1 transgenic mice, also known as beta amyloid precursor protein / The spatial memory function of presenilin 1 transgenic mice or Alzheimer's model mice improves their cognitive function decline.

Specifically, the elderly APP/PS1 transgenic mice were intraperitoneally injected with different concentrations of FG-4592, and the spatial memory function and the key enzyme of APP-cleaving BACE1 (also known as β-site secretion) were detected after 2-3 weeks of continuous administration. Expression of enzyme 1). It was found that different concentrations of FG-4592 in APP/PS1 transgenic mice can improve the spatial memory function of mice, down-regulate the expression of BACE1, a key enzyme of splicing APP, and the effect of low-dose long-term administration is short. Dosing is more effective. Therefore, using FG-4592 and its pharmaceutically acceptable salt as a medicinal ingredient or active ingredient for improving cognitive function of human or animal, and improving memory, only a small dose can be administered for a period of time without continuous administration. A larger dose is required.

DRAWINGS

Figure 1 is a flow chart for detecting the spatial memory function of mice using the Morris water maze method.

Figure 2 is a graph showing the effect of different doses of FG-4592 on learning ability of mice; wherein A is: the effect of FG-4592 of 30 mg/kg on the learning ability of mice; B is: the pair of FG-4592 of 15 mg/kg The impact map of mouse learning ability.

Figure 3 is a graph of the number of mouse platform crossings administered to different doses of FG-4592; wherein A is: a map of the number of mouse platform crossings of FG-4592 administered with 30 mg/kg; B is: FG-4592 administered 15 mg/kg Mouse platform crossing times map.

Figure 4 is a plot of the latency of the first passage of the platform to mice administered different doses of FG-4592; wherein A is: the latency map of the first time the mouse is administered 30 mg/kg of FG-4592 across the platform; B is: 15 mg/kg administered The FG-4592 mouse first crossed the platform for the incubation period.

Figure 5 is a route of movement of mice administered different doses of FG-4592 in the target quadrant; wherein A is: the distance traveled by the mice given 30 mg/kg of FG-4592 in the target quadrant; B is: 15 mg/kg administered The FG-4592 mouse travels in the target quadrant.

Figure 6 is a time-series diagram of mice administered different doses of FG-4592 in the target quadrant; wherein A is: a time course map of the mouse given 30 mg/kg of FG-4592 in the target quadrant; B is: administered Time course plot of 15 mg/kg of FG-4592 mice in the target quadrant.

Figure 7 is a typical pathway diagram of the effects of spatial memory on mice administered different doses of FG-4592; wherein A is: a typical pathway map of the effect of spatial memory on mice given 30 mg/kg of FG-4592; B is: administration A typical path map of the spatial memory effects of FG-4592 in mice at 15 mg/kg (n=10, *: p<0.05).

Figure 8 is a graph showing the effect of FG-4592 on the expression of HIF-1α, BACE1 and APP-related proteins in mouse brain tissue; wherein A is: FG-4592 can up-regulate HIF-1α and down-regulate BACE1 expression; B is :FG-4592 regulates the expression of HIF-1α; C is: the effect of FG-4592 on the full-length expression of APP; D is: the effect of FG-4592 on the expression of C-terminal fragment of APP; E is: FG-4592 The expression pattern of BACE1 was adjusted (n=4, *: p<0.05).

Detailed ways

Preferred embodiments of the present invention are described in detail below. The preferred embodiments described herein are to be construed as illustrative only and not limiting.

The experimental methods described in the following examples are conventional methods unless otherwise specified; the reagents and materials are commercially available unless otherwise specified.

The FG-4592 used in the following examples was purchased from Selleck, Inc., USA, catalog number: S1007, batch: 03, as a white powder.

Example 1

Protective effect of FG-4592 on cognitive function in aged APP/PS1 transgenic mice

1.FG-4592 drug and mouse treatment

(1) Treatment of FG-4592 drug: 100 mg of FG-4592 whole drug was dissolved in 22 ml and 11 ml of 5% (volume ratio) DMSO + 50% (volume ratio) PEG300 + ddH2O solution; Store at -20 ° C. Wherein DMSO is dimethyl sulfoxide; PEG 300 is polyethylene glycol 300; ddH2O means double distilled water, that is, water obtained by two distillations.

(2) Treatment of mice: The experimental mice were 6-month-old APP/PS1 transgenic mice weighing 30±5 g, male. During the experiment, the animals were kept in a standardized animal house and were free to eat and drink. According to the body weight of the mice, the mice were intraperitoneally injected with the FG-4592 solution prepared in the step (1) once every other day, each injection volume was 100 μl. When the total injection dose was 15 mg/kg, the administration time lasted for 3 weeks; when the total injection dose was 30 mg/kg, the administration time lasted for 2 weeks.

(3) Control group: APP/PS1 transgenic mice were not given drug treatment with FG-4592, and the memory level of the hidden platform was measured by the normal aged mice measured by the water maze test.

2. Experimental methods

Morris water maze experiments were divided into learning evaluation and memory function evaluation. The water maze is evenly divided into 4 quadrants. During the study, the mice were placed in a water maze every day to train them to find a transparent platform hidden 1.5 cm below the surface of the water, once per quadrant per day, and to record the time of each stage of the mouse. When the memory function is evaluated, the platform hidden under the water surface is taken out, and the mouse swims freely for 1 min in the water maze. The movement trajectory of the mouse, the time of crossing the platform for the first time, the number of crossing platforms, the movement time in the target quadrant, and the target are recorded. The distance of movement in the quadrant.

In the Western Blotting experiment, the animal brain tissue was taken out by conventional method, and the protein was homogenized to obtain Western blotting. The changes of APP cleavage and the expression of key enzyme BACE1 were detected. The results obtained were evaluated for differences between groups using an independent sample t test or a One-way ANOVA method.

3. Experimental results

The spatial memory function of mice was detected by Morris water maze method. The dose schedule of intraperitoneal injection of FG-4592 in mice and the detection scheme of Morris water maze behavior are shown in Figure 1. The results showed that in the training, administration of the 30 mg/kg dose of FG-4592 (dose cycle of 2 weeks) had no significant effect on the learning ability of the mice (see Figure 2A); while the FG was given at a dose of 15 mg/kg. -4592 (dose cycle of 3 weeks) can significantly improve the learning ability of mice. The mice started on the fifth day of learning, and the time of landing on the platform was significantly reduced compared with the control group. The mice were in the Morris water maze. The incubation period for finding hidden platforms during training was significantly shorter than for control mice (see Figure 2B).

Figure 7 is a typical pathway map of spatial memory testing in mice after administration of different doses of FG-4592. The results showed that administration of a 30 mg/kg dose of FG-4592 (dose cycle of 2 weeks) significantly increased the number of times the mouse crossed the platform (see Figure 3A), shortening the latency of the mouse crossing the platform for the first time (see Figure 4A). However, there is no significant improvement in the travel distance of the target quadrant (see Figure 5A) and the time spent in the target quadrant (see Figure 6A). Administration of FG-4592 at a dose of 15 mg/kg (dose cycle of 3 weeks) can significantly reduce the latency of the first crossing of the platform (see Figure 4B), increasing the number of crossings (see Figure 3B) and the distance traveled in the target quadrant (See Figure 5B), and the time of stay in the target quadrant (see Figure 6B). This result indicates that intraperitoneal injection of FG-4592 in APP/PS1 mice can significantly improve the spatial memory ability of mice and improve their cognitive function.

The results of Western Blotting are shown in Figure 8, where the control group is the normal protein expression level of the mice that are not given drug treatment. The results included: intraperitoneal injection of FG-4592 down-regulated BACE1 expression by stabilizing HIF-1α (Fig. 8A); administration of FG-4592 increased HIF-1α expression in mouse brain tissue (see Figure 8B), for APP Both the long and C-terminal fragments had no significant effect (see Figure 8C, Figure 8D), but the expression of the key enzyme BACE1, which is APP-cleaved, was down-regulated (see Figure 8E). This result indicates that down-regulation of BACE1 plays an important role in improving cognitive function in mice by FG-4592.

Among them, in Figure 8:

The APP-Ful Chinese name is: the full length of the beta amyloid precursor protein;

The English name of APP-CTF is APP-Cleaved carboxy-terminal fragment, and the Chinese name is: β-amyloid precursor protein carbon-terminal cleavage fragment;

The β-actin Chinese name is: actin.

Note: In Figure 7 and Figure 8, n indicates the number of experimental animals in each group in the experiment, *: p < 0.05 indicates that the data obtained in the experiment is statistically significant.

The foregoing description of the specific exemplary embodiments of the present invention has The description is not intended to limit the invention to the precise forms disclosed. The embodiments were chosen and described in order to explain the particular embodiments of the invention Choose and change. The scope of the invention is intended to be defined by the claims and their equivalents.

Claims (10)

Use of FG-4592, or a pharmaceutically acceptable salt thereof, for the manufacture of a medicament for the treatment and/or prevention of a neurological or neurodegenerative disease. The use according to claim 1, wherein the nerve damage disease is early cognitive impairment, stroke; the neurodegenerative disease is Alzheimer's disease, cognitive dysfunction, vascular dementia, Pa One or more of Jinsen's disease. The use according to claim 1, characterized in that the product is a medicine and/or a health product. The use according to claim 1, characterized in that the product is capable of improving the spatial memory function of a human or an animal. A medicine for treating and/or preventing a nerve damage disease or a neurodegenerative disease, wherein the active ingredient is FG-4592 or a pharmaceutically acceptable salt thereof. The pharmaceutical product according to claim 5, wherein the pharmaceutical product further comprises one or more pharmaceutically acceptable excipients or carriers. The medicine according to claim 5, wherein the pharmaceutical preparation is prepared from a tablet, a capsule, an effervescent tablet, a granule, a powder, a dispersion tablet, an oral solution, a pill or an injection. A health care product for treating and/or preventing a nerve damage disease or a neurodegenerative disease, wherein the active ingredient is FG-4592 or a pharmaceutically acceptable salt thereof. The health care product according to claim 8, wherein the health care product further comprises one or more excipients or carriers adapted to different dosage forms. The health care product according to claim 8, wherein the health care product is formulated into a tablet, a capsule or an oral solution.

Images