KR20240045472A - Composition for treating osteoarthritis comprising quinazoline salt compound and preparing method of the same - Google Patents

Abstract

The present application relates to a composition for treating arthritis, comprising a quinazoline salt compound represented by the following formula (1):
[Formula 1]

(In Formula 1 above,
A is HCl, HClO ₃ , H ₂ SO ₄ , HNO ₃ , HI or HBr,
R ₁ , R ₂ , and R ₃ are each independently optionally substituted linear or branched C ₁ -C ₁₂ alkyl, optionally substituted C ₃ -C ₁₂ cycloalkyl, optionally substituted C ₆ -C ₁₈ Aryl, or alkoxy,
The substitution is by oxygen, nitrogen, sulfur, linear or branched C ₁ -C ₆ alkyl, C ₆ -C ₂₀ aryl, halogen, alkoxy, trimethylsilyl, ether, or a combination thereof) .

Description

Composition for treating arthritis comprising a quinazoline salt compound and method for manufacturing the same {COMPOSITION FOR TREATING OSTEOARTHRITIS COMPRISING QUINAZOLINE SALT COMPOUND AND PREPARING METHOD OF THE SAME}

The present application relates to a composition for treating arthritis containing a quinazoline salt compound and a method for producing the same.

Arthritis is an inflammatory disease of the joints, and the diseases with the largest number of cases are chronic rheumatoid arthritis and deforming arthrosis (osteoarthrosis).

Chronic joint rheumatism is thought to be an autoimmune disease and is accompanied by symptoms of joint pain, stiffness, and swelling. However, as the condition progresses, it often leads to degeneration of the articular cartilage surface such as deforming arthrosis. , and eventually severe destruction of the bones and cartilage of the joint occurs.

Arthropathy deformity is a degenerative disease of the articular cartilage that frequently occurs in elderly people. Osteoarthritis (OA) is a disease that causes destruction of cartilage and proliferative changes in bone or cartilage due to degenerative degeneration of joint components, and these changes can also cause secondary arthritis (e.g. , synovitis). Degenerative arthrosis occurs frequently in load joints such as the knee joint, elbow joint, and hip joint, and is also confirmed to occur in non-load joints such as the shoulder joint, elbow joint, and hand joint. Also, temporomandibular arthropathy, which presents the same condition in the temporomandibular joint, is known.

Currently, the standard treatments for osteoarthritis that are widely used include anti-inflammatory analgesics administered orally, hyaluronic acid or corticosteroids injected into joints, which are all inhibitors of joint pain and do not cause osteoarthritis. There is a need for the development of pharmaceutical compositions that reduce gene expression.

Republic of Korea Patent No. 10-1207155 relates to a treatment or prevention agent for arthritis. The patent discloses a treatment for arthritis containing C-type natriuretic peptide (CNP) or a derivative thereof as an active ingredient, but does not mention a composition for the treatment of arthritis containing a quinazoline salt compound.

The present application aims to solve the problems of the prior art described above and to provide a composition for treating arthritis containing a quinazoline salt compound.

Additionally, the object is to provide a method for producing the composition for treating arthritis.

Another object is to provide an intra-articular injection containing the composition for treating arthritis.

However, the technical challenges sought to be achieved by the embodiments of the present application are not limited to the technical challenges described above, and other technical challenges may exist.

As a technical means for achieving the above technical problem, the first aspect of the present application provides a composition for treating arthritis, comprising a quinazoline salt compound represented by the following formula (1):

[Formula 1]

(In Formula 1 above,

A is HCl, HClO ₃ , H ₂ SO ₄ , HNO ₃ , HI or HBr,

R ₁ , R ₂ , and R ₃ are each independently optionally substituted linear or branched C ₁ -C ₁₂ alkyl, optionally substituted C ₃ -C ₁₂ cycloalkyl, optionally substituted C ₆ -C ₁₈ Aryl, or alkoxy,

The substitution is by oxygen, nitrogen, sulfur, linear or branched C ₁ -C ₆ alkyl, C ₆ -C ₂₀ aryl, halogen, alkoxy, trimethylsilyl, ether, or a combination thereof) .

According to one embodiment of the present application, the quinazoline salt may include the following compounds, but is not limited thereto:

.

According to one embodiment of the present application, the arthritis may include, but is not limited to, degenerative arthritis.

In addition, the second aspect of the present application includes preparing a quinazoline derivative by reacting an aromatic aldehyde and a carbamate; and reacting the quinazoline derivative with an acid to produce a quinazoline salt.

According to one embodiment of the present application, the aromatic aldehyde may include, but is not limited to, 4-hydroxy-3,5-dimethyl benzaldehyde.

According to one embodiment of the present application, the carbamate may include Tert-Butyl N-(2-bromoethyl)carbamate, but is limited thereto. That is not the case.

According to one embodiment of the present application, the quinazoline derivative may include a compound represented by the following formula (2), but is not limited thereto:

[Formula 2]

(In Formula 2 above,

R ₁ , R ₂ , and R ₃ are each independently optionally substituted linear or branched C ₁ -C ₁₂ alkyl, optionally substituted C ₃ -C ₁₂ cycloalkyl, optionally substituted C ₆ -C ₁₈ Aryl, or alkoxy,

The substitution is by oxygen, nitrogen, sulfur, linear or branched C ₁ -C ₆ alkyl, C ₆ -C ₂₀ aryl, halogen, alkoxy, trimethylsilyl, ether, or a combination thereof) .

According to one embodiment of the present application, the acid may include, but is not limited to, one selected from the group consisting of HCl, HClO ₃ , H ₂ SO ₄ , HNO ₃ , HI, HBr, and combinations thereof.

According to one embodiment of the present application, the quinazoline salt may include a compound represented by the following formula (3), but is not limited thereto:

[Formula 3]

(In Formula 3 above,

A is HCl, HClO ₃ , H ₂ SO ₄ , HNO ₃ , HI or HBr,

R ₁ , R ₂ , and R ₃ are each independently optionally substituted linear or branched C ₁ -C ₁₂ alkyl, optionally substituted C ₃ -C ₁₂ cycloalkyl, optionally substituted C ₆ -C ₁₈ Aryl, or alkoxy,

The substitution is by oxygen, nitrogen, sulfur, linear or branched C ₁ -C ₆ alkyl, C ₆ -C ₂₀ aryl, halogen, alkoxy, trimethylsilyl, ether, or a combination thereof) .

According to one embodiment of the present application, the quinazoline salt may include the following compounds, but is not limited thereto:

.

Additionally, a third aspect of the present application provides an intra-articular injection comprising the composition for treating arthritis according to the first aspect of the present application.

The above-described means of solving the problem are merely illustrative and should not be construed as intended to limit the present application. In addition to the exemplary embodiments described above, additional embodiments may be present in the drawings and detailed description of the invention.

The composition for treating arthritis according to the present application includes a quinazoline salt compound, a substance that has not previously been reported for use in treating inflammation. Specifically, the composition for treating arthritis may have an effect of suppressing osteoarthritis by reducing the expression of genes that cause osteoarthritis, and may be particularly effective in treating degenerative arthritis.

However, the effects that can be obtained herein are not limited to the effects described above, and other effects may exist.

1 is a flowchart of a method for producing a composition for treating arthritis according to an embodiment of the present application.
Figure 2 shows NMR results of a quinazoline salt compound prepared according to an example of the present application.
Figure 3 shows LC-MS results of a quinazoline salt compound prepared according to an example of the present application.
Figure 4 shows the results of PCR performed to confirm the anti-inflammatory effect of a composition for treating arthritis according to an experimental example herein.
Figure 5 shows the results of qRT-PCR performed to confirm the anti-inflammatory effect of a composition for treating arthritis according to an experimental example herein.
Figure 6 shows the results of a Western blot performed to confirm the anti-inflammatory effect of a composition for treating arthritis according to an experimental example herein.
Figure 7 shows the results of Safranin-O staining according to an experimental example herein.
Figure 8 shows the results of the histopathological evaluation method proposed by OARSI (Osteoarthritis Research Society International) according to an experimental example of the present institution.

Below, with reference to the attached drawings, embodiments of the present application will be described in detail so that those skilled in the art can easily implement them. However, the present application may be implemented in various different forms and is not limited to the embodiments described herein. In order to clearly explain the present application in the drawings, parts that are not related to the description are omitted, and similar reference numerals are assigned to similar parts throughout the specification.

Throughout this specification, when a part is said to be “connected” to another part, this includes not only the case where it is “directly connected,” but also the case where it is “electrically connected” with another element in between. do.

Throughout this specification, when a member is said to be located “on”, “above”, “at the top”, “below”, “at the bottom”, or “at the bottom” of another member, this means that a member is located on another member. This includes not only cases where they are in contact, but also cases where another member exists between two members.

Throughout the specification of the present application, when a part is said to “include” a certain element, this means that it may further include other elements rather than excluding other elements, unless specifically stated to the contrary.

As used herein, the terms “about,” “substantially,” and the like are used to mean at or close to a numerical value when manufacturing and material tolerances inherent in the stated meaning are presented, and to aid understanding of the present application. It is used to prevent unscrupulous infringers from unfairly exploiting disclosures in which precise or absolute figures are mentioned. Additionally, throughout the specification herein, “a step of” or “a step of” does not mean “a step for.”

Throughout this specification, the term "combination thereof" included in the Markushi format expression means a mixture or combination of one or more components selected from the group consisting of the components described in the Markushi format expression, It means including one or more selected from the group consisting of.

Throughout this specification, description of “A and/or B” means “A, B, or A and B.”

Hereinafter, the composition for treating arthritis containing the quinazoline salt compound of the present application and the method for manufacturing the same will be described in detail with reference to embodiments, examples, and drawings. However, the present application is not limited to these embodiments, examples, and drawings.

As a technical means for achieving the above technical problem, the first aspect of the present application provides a composition for treating arthritis, comprising a quinazoline salt compound represented by the following formula (1):

[Formula 1]

(In Formula 1 above,

A is HCl, HClO ₃ , H ₂ SO ₄ , HNO ₃ , HI or HBr,

R ₁ , R ₂ , and R ₃ are each independently optionally substituted linear or branched C ₁ -C ₁₂ alkyl, optionally substituted C ₃ -C ₁₂ cycloalkyl, optionally substituted C ₆ -C ₁₈ Aryl, or alkoxy,

The substitution is by oxygen, nitrogen, sulfur, linear or branched C ₁ -C ₆ alkyl, C ₆ -C ₂₀ aryl, halogen, alkoxy, trimethylsilyl, ether, or a combination thereof) .

The composition for treating arthritis according to the present application includes a quinazoline salt compound, a substance that has not previously been reported for use in treating inflammation. Specifically, the composition for treating arthritis may have an effect of suppressing osteoarthritis by reducing the expression of genes that cause osteoarthritis, and may be particularly effective in treating degenerative arthritis.

According to one embodiment of the present application, the quinazoline salt may include the following compounds, but is not limited thereto:

.

The compound is 2-[4-[2-(dimethylamino)ethoxy]-3,5-dimethyl-phenyl]-5,7-dimethoxy-3H-quinazolin-4-one, dihydrochloride [2- [4-[2-(dimethylamino)ethoxy]-3,5-dimethyl-phenyl]-5,7-dimethoxy-3H-quinazolin-4-one, dihydrochloride], which has not been previously used as a composition for treating arthritis. material, and the inventors of the present application discovered a novel use of the quinazoline salt compound as a composition for treating arthritis.

According to one embodiment of the present application, the arthritis may include, but is not limited to, degenerative arthritis.

In addition, the second aspect of the present application includes preparing a quinazoline derivative by reacting an aromatic aldehyde and a carbamate; and reacting the quinazoline derivative with an acid to produce a quinazoline salt.

Regarding the method for producing a composition for treating arthritis according to the second aspect of the present application, detailed description of parts overlapping with the first aspect of the present application has been omitted. However, even if the description is omitted, the information described in the first aspect of the present application is the same as the present application. The same can be applied to the second aspect of .

1 is a flowchart of a method for producing a composition for treating arthritis according to an embodiment of the present application.

First, a quinazoline derivative is prepared by reacting aromatic aldehyde and carbamate (S100).

According to one embodiment of the present application, the aromatic aldehyde may include, but is not limited to, 4-hydroxy-3,5-dimethyl benzaldehyde.

According to one embodiment of the present application, the carbamate may include Tert-Butyl N-(2-bromoethyl)carbamate, but is limited thereto. That is not the case.

According to one embodiment of the present application, the quinazoline derivative may include a compound represented by the following formula (2), but is not limited thereto:

[Formula 2]

(In Formula 2 above,

R ₁ , R ₂ , and R ₃ are each independently optionally substituted linear or branched C ₁ -C ₁₂ alkyl, optionally substituted C ₃ -C ₁₂ cycloalkyl, optionally substituted C ₆ -C ₁₈ Aryl, or alkoxy,

The substitution is by oxygen, nitrogen, sulfur, linear or branched C ₁ -C ₆ alkyl, C ₆ -C ₂₀ aryl, halogen, alkoxy, trimethylsilyl, ether, or a combination thereof) .

For example, the quinazoline derivative may include, but is not limited to, the one represented by Compound 1 below:

[Compound 1]

.

In Compound 1, the protecting group Boc group (tert-butylcarbonyl group) is a material selected from the group consisting of trifluoroacetic acid (TFA), hydrochloric acid, hydrogen chloride, trimethylsilyl iodide, aluminum chloride, and combinations thereof. It may be deprotected and substituted with hydrogen and have an amine group at the end, but is not limited thereto. For example, Compound 1 whose terminal is substituted with an amine group may include, but is not limited to, Compound 2 below.

[Compound 2]

.

In Compound 2, the hydrogen of the amine group may be replaced with an alkyl group by a substance that acts as a strong reducing agent such as formaldehyde, sodium borohydride, etc., but is not limited thereto. For example, the compound in which the hydrogen of the amine group of Compound 2 is replaced with an alkyl group may include, but is not limited to, Compound 3 below.

[Compound 3]

.

Next, the quinazoline derivative is reacted with acid to prepare a quinazoline salt (S200).

According to one embodiment of the present application, the acid may include, but is not limited to, one selected from the group consisting of HCl, HClO ₃ , H ₂ SO ₄ , HNO ₃ , HI, HBr, and combinations thereof.

According to one embodiment of the present application, the quinazoline salt may include a compound represented by the following formula (3), but is not limited thereto:

[Formula 3]

(In Formula 3 above,

A is HCl, HClO ₃ , H ₂ SO ₄ , HNO ₃ , HI or HBr,

R ₁ , R ₂ , and R ₃ are each independently optionally substituted linear or branched C ₁ -C ₁₂ alkyl, optionally substituted C ₃ -C ₁₂ cycloalkyl, optionally substituted C ₆ -C ₁₈ Aryl, or alkoxy,

The substitution is by oxygen, nitrogen, sulfur, linear or branched C ₁ -C ₆ alkyl, C ₆ -C ₂₀ aryl, halogen, alkoxy, trimethylsilyl, ether, or a combination thereof) .

According to one embodiment of the present application, the quinazoline salt may include the following compounds, but is not limited thereto:

.

Additionally, a third aspect of the present application provides an intra-articular injection comprising the composition for treating arthritis according to the first aspect of the present application.

Regarding the intra-articular injection according to the third aspect of the present application, detailed description of parts overlapping with the first aspect of the present application has been omitted. However, even if the description is omitted, the contents described in the first aspect of the present application are the third aspect of the present application. The same can be applied to .

An intra-articular injection containing the composition for treating arthritis according to the present application may be administered into the joint space to reduce the expression of genes that cause osteoarthritis and thereby have an effect of suppressing osteoarthritis.

The present invention will be described in more detail through the following examples. However, the following examples are for illustrative purposes only and are not intended to limit the scope of the present application.

[Example 1] Preparation of composition for treating arthritis

First, a quinazoline salt compound was prepared according to Scheme 1 below.

[Scheme 1]

The prepared quinazoline salt compound was confirmed by NMR, and its purity was confirmed by LC-MS.

Figure 2 shows NMR results of a quinazoline salt compound prepared according to an example of the present application.

Figure 3 shows LC-MS results of a quinazoline salt compound prepared according to an example of the present application.

2 and 3, it was confirmed that the quinazoline salt compound according to an example of the present application was actually produced. Next, the prepared quinazoline salt compound was dissolved in dimethyl sulfoxide (DMSO) to prepare a composition for treating arthritis. Manufactured.

[Comparative example]

As a comparative example, JQ-1, which is used as a BET (Bromodomain and extra terminal containing protein) inhibitor, was used.

[Experimental Example 1] In vitro anti-inflammatory efficacy test

An experiment was performed to confirm the anti-inflammatory effect of the composition for treating arthritis according to an example herein.

First, cartilage cells were isolated from mouse joints using collagenase.

Subsequently, the separated chondrocytes were cultured in FBS/DMEM medium for 3 days and treated with interleukin 1-beta (IL-1β) to induce inflammation, and then the composition for treating arthritis of the examples was added at different concentrations (0 μM to 20 μM). ) was treated for 24 hours.

Subsequently, RNA was extracted using Trizol, and the pattern of Mmp3, Mmp13, Cox2, and IL-6 gene expression by treatment with the composition for treating arthritis of the example was confirmed through PCR, qRT-PCR, and western blot.

Figure 4 shows the results of PCR performed to confirm the anti-inflammatory effect of a composition for treating arthritis according to an experimental example herein.

Figure 5 shows the results of qRT-PCR performed to confirm the anti-inflammatory effect of a composition for treating arthritis according to an experimental example herein.

Figure 6 shows the results of a Western blot performed to confirm the anti-inflammatory effect of a composition for treating arthritis according to an experimental example herein.

Referring to Figures 4 to 6, it was confirmed that as the treatment concentration of the examples increased, the expression of Mmp3, Mmp13, Cox2, and IL-6 genes tended to decrease. In addition, a decrease in the expression of genes causing osteoarthritis was observed compared to the comparative example (JQ-1) used as a control. In conclusion, the above test results confirmed the osteoarthritis inhibition effect of the test substance (Example 1).

[Experimental Example 2] Efficacy test using degenerative arthritis animal model (DMM)

A 12-week-old B6 mouse was anesthetized, the knee was cut in a straight line using a No. 11 blade, the knee ligament was pushed to the side, and the meniscus was identified, excised, and sutured. On the other leg, a portion of the knee skin was excised in a straight line, and the knee ligament was pushed to the side and restored to its original state. Degenerative arthritis was induced, and after 4 weeks, the test substance (Example 1) was injected intraarticularly into the mouse knee joint at different doses (25, 50, and 100 mg/kg). Six weeks after starting test substance administration, mouse knees were fixed with 4% PFA and decalcification was performed for 2 weeks using EDTA (ethylene-diamine-tetraacetic acid) solution. Paraffin was absorbed into knee cartilage that had undergone a paraffinization process, and then Safranin-O staining was performed to confirm bone tissue.

Figure 7 shows the results of Safranin-O staining according to an experimental example herein.

Referring to Figure 7, after inducing degenerative arthritis (DMM) in B6 mice, test substances (Example 1) were administered at different doses, and bone tissue was confirmed through Safranin O-staining at 6 weeks. As a result, it was confirmed that the effect of inhibiting cartilage destruction was shown in a dose-dependent manner.

In addition, the histopathological evaluation method suggested by OARSI (Osteoarthritis Research Society International) was performed to confirm the ability of the composition for treating arthritis according to Example 1 herein to inhibit destruction of mouse knee cartilage. The OARSI evaluation method is an evaluation method that gives scores ranging from 0 to 6 depending on the degree of cartilage damage, and the scoring criteria are shown in Table 1 below. Additionally, since SBP (subchondral bone plate) thickness increases when osteoarthritis occurs, we also checked whether SBP thickness changed when drug was administered.

[Table 1] Scoring criteria according to the OARSI evaluation method

Figure 8 shows the results of performing the histopathological evaluation method proposed by OARSI according to an experimental example of the present institution.

Figure 9 shows the results of confirming the change in SBP thickness according to an experimental example herein.

Referring to Figures 8 and 9, it was confirmed that as the administered dose of the test substance increased, the OARSI grade decreased and the SBP thickness decreased. This confirmed that the test substance had a dose-dependent osteoarthritis treatment effect.

The description of the present application described above is for illustrative purposes, and those skilled in the art will understand that the present application can be easily modified into other specific forms without changing its technical idea or essential features. Therefore, the embodiments described above should be understood in all respects as illustrative and not restrictive. For example, each component described as unitary may be implemented in a distributed manner, and similarly, components described as distributed may also be implemented in a combined form.

The scope of the present application is indicated by the claims described below rather than the detailed description above, and all changes or modified forms derived from the meaning and scope of the claims and their equivalent concepts should be construed as being included in the scope of the present application.

Claims (11)

A composition for treating arthritis, comprising a quinazoline salt compound represented by the following formula (1):
[Formula 1]

(In Formula 1 above,
A is HCl, HClO ₃ , H ₂ SO ₄ , HNO ₃ , HI or HBr,
R ₁ , R ₂ , and R ₃ are each independently optionally substituted linear or branched C ₁ -C ₁₂ alkyl, optionally substituted C ₃ -C ₁₂ cycloalkyl, optionally substituted C ₆ -C ₁₈ Aryl, or alkoxy,
The substitution is by oxygen, nitrogen, sulfur, linear or branched C ₁ -C ₆ alkyl, C ₆ -C ₂₀ aryl, halogen, alkoxy, trimethylsilyl, ether, or a combination thereof) .
According to claim 1,
A composition for treating arthritis, wherein the quinazoline salt includes the following compounds:
.
According to claim 1,
The arthritis includes degenerative arthritis,
Composition for treating arthritis.
Preparing a quinazoline derivative by reacting aromatic aldehyde and carbamate; and
Method for producing a composition for treating arthritis, comprising the step of reacting the quinazoline derivative with an acid to produce a quinazoline salt:
According to claim 4,
A method of producing a composition for treating arthritis, wherein the aromatic aldehyde includes 4-hydroxy-3,5-dimethyl benzaldehyde.
According to claim 4,
A method of producing a composition for treating arthritis, wherein the carbamate includes Tert-Butyl N-(2-bromoethyl)carbamate.
According to claim 4,
A method for producing a composition for treating arthritis, wherein the quinazoline derivative includes a compound represented by the following formula (2):
[Formula 2]

(In Formula 2 above,
R ₁ , R ₂ , and R ₃ are each independently optionally substituted linear or branched C ₁ -C ₁₂ alkyl, optionally substituted C ₃ -C ₁₂ cycloalkyl, optionally substituted C ₆ -C ₁₈ Aryl, or alkoxy,
The substitution is by oxygen, nitrogen, sulfur, linear or branched C ₁ -C ₆ alkyl, C ₆ -C ₂₀ aryl, halogen, alkoxy, trimethylsilyl, ether, or a combination thereof) .
According to claim 4,
The acid includes one selected from the group consisting of HCl, HClO ₃ , H ₂ SO ₄ , HNO ₃ , HI, HBr and combinations thereof,
Method for producing a composition for treating arthritis.
According to claim 4,
A method for producing a composition for treating arthritis, wherein the quinazoline salt includes a compound represented by the following formula (3):
[Formula 3]

(In Formula 3 above,
A is HCl, HClO ₃ , H ₂ SO ₄ , HNO ₃ , HI or HBr,
R ₁ , R ₂ , and R ₃ are each independently optionally substituted linear or branched C ₁ -C ₁₂ alkyl, optionally substituted C ₃ -C ₁₂ cycloalkyl, optionally substituted C ₆ -C ₁₈ Aryl, or alkoxy,
The substitution is by oxygen, nitrogen, sulfur, linear or branched C ₁ -C ₆ alkyl, C ₆ -C ₂₀ aryl, halogen, alkoxy, trimethylsilyl, ether, or a combination thereof) .
According to clause 9,
A method for producing a composition for treating arthritis, wherein the quinazoline salt includes the following compounds:
.
An intraarticular injection comprising the composition for treating arthritis according to any one of claims 1 to 3.