Methods for treating rheumatoid arthritis (RA) using enzyme- and substrate-selective exosite inhibitors of a disintegrin and metalloprotease 10 (ADAM10)

Abstract

The present invention provides methods for treating immune dysfunction conditions, such as rheumatoid arthritis (RA), using an enzyme- and substrate-selective exosite inhibitor of a disintegrin and metalloprotease 10 (ADAM10), such as N-(3-chloro-4-methylphenyl)-2-[2-(3-methoxybenzoyl)hydrazinyl]-2-oxoacetamide (PubChem CID3117694). A method for treating an immune dysfunction condition in a subject in need thereof includes providing a composition comprising a therapeutically effective dose of a modulator of a disintegrin and metalloprotease 10 (ADAM10) and at least one acceptable pharmaceutical carrier, and administering the composition to the subject to treat the immune dysfunction condition in the subject through modulation of ADAM10. A composition comprising a therapeutically effective dose of a modulator of a disintegrin and metalloprotease 10 (ADAM10), a therapeutically effective dose of indomethacin, and at least one acceptable pharmaceutical carrier, is used to treat a condition of immune dysfunction or inflammation in a subject in need thereof.

Description

[Technical field]

[0001] The present invention is encompassed in the field of therapeutic agents for inflammatory and / or autoimmune conditions, and relates generally to the treatment of rheumatoid arthritis (RA), and in particular to the use of inhibitors of a disintegrin and metalloprotease 10 (ADAM10) for the treatment of RA, and in particular to the use of N-(3-chloro-4-methylphenyl)-2-[2-(3-methoxybenzoyl)hydrazinyl]-2-oxoacetamide (PubChem CID3117694) for the treatment of RA. [Background technology]

[0002] A disintegrin and metaprotease 10 (ADAM10) is a member of a large group of human and non-human zinc-dependent enzymes (reviewed in Cerda-Costa and Gomis-Ruth, 2014). Structurally, it belongs to the adamalysin family (Minond, 2020, ADAM and ADAMTS enzymes). ADAM10 is a cell surface enzyme that cleaves a wide variety of cell surface proteins (Dreymueller et al., 2015; Kuhn et al., 2016; Camodeca et al., 2019; Scharfenberg et al., 2019), and has been suggested to have importance in cancer progression, inflammation, and immune responses and may be an important target for therapy, for example, but not limited to, the treatment of rheumatoid arthritis (RA).

[0003] Rheumatoid arthritis is a common disease that affects millions of people in the United States alone. It is a chronic inflammatory autoimmune condition that affects the entire body and can cause permanent damage to joints. There is no known cure.

[0004] Disease-modifying antirheumatic drugs (DMARDs) approved for the treatment of RA include agents with the following mechanisms of action: Janus kinase (JAK) inhibition, TNF inhibition, T cell costimulation blockade, IL-6 receptor inhibition, B cell depletion, and interleukin-1 inhibition. Although ACR70 (a DMARD) has shown improved efficacy over methotrexate monotherapy (70% improvement in responders), the response rate is less than 50%. Thus, there remains an unmet need for antirheumatic drugs that improve the efficacy and response rate of ACR70, either alone or in combination with approved drugs. Summary of the Invention [Problem to be solved by the invention]

[0005] The invention described herein provides novel selective ADAM10 inhibitors that fulfill this unmet need for improved antirheumatic drugs. These novel selective ADAM10 inhibitors act via a non-Zn-binding mechanism and potentially bind to a previously unknown secondary substrate-binding site (exosite) outside the active site.

[0006] In one basic aspect, the present invention provides a new treatment approach for inflammation and / or immune dysfunction. Inflammation is considered to be the response of tissues to infection or injury, resulting in tissue swelling, redness, heat, and / or pain. Immune dysfunction is considered to be any activity or function of the immune system that differs from healthy function and leads to undesirable consequences. [Means for solving the problem]

[0007] In one basic aspect, the present invention provides new treatments for rheumatoid arthritis (RA). In another basic aspect, the present invention provides novel compositions and treatments for rheumatoid arthritis.

[0008] In one aspect, the present invention provides a pharmaceutical composition for treating immune dysfunction, autoimmunity, inflammation, and / or rheumatoid arthritis, comprising a therapeutically effective dose of a modulator of a disintegrin and metalloprotease 10 (ADAM10) and at least one acceptable pharmaceutical carrier. An "acceptable pharmaceutical carrier" can be any inert and non-toxic agent useful in the preparation of a medicament. The phrase "therapeutically effective dose" or "therapeutically effective amount" refers to the amount of a composition required to achieve a desired function, e.g., alleviation of symptoms of inflammation, immune dysfunction, and / or rheumatoid arthritis. A "modulator" can refer to any composition, compound, or substance that causes a change in the function and / or activity of another composition, compound, or substance. A non-limiting example of a modulator is a protein or enzyme inhibitor. A protein / enzyme inhibitor reduces and / or stops the activity and / or function of a protein or enzyme. A specific non-limiting example is an inhibitor of a disintegrin and metalloprotease 10 (ADAM10). The term "autoimmunity" refers to an immune response of an organism against its own healthy cells and / or tissues. Pharmaceutical compositions can be formulated for a particular situation, for example, for oral, sublingual, buccal, nasal, rectal, parenteral, intradermal, transdermal, or intratracheal administration.

[0009] In another aspect, the present invention provides various methods of treatment for a subject in need of treatment. These various methods include, but are not limited to, treating immune dysfunction, treating autoimmunity, reducing inflammation, and / or treating rheumatoid arthritis. The term "subject" refers to any human or animal that will benefit from the use of the compositions, methods, and / or treatments described herein. A preferred, but non-limiting, subject is a human patient with rheumatoid arthritis. The method includes providing a composition comprising a therapeutically effective dose of a modulator of a disintegrin and metalloprotease 10 (ADAM10) and an acceptable pharmaceutical carrier, and administering the composition to the subject. The modulator can be, but is not limited to, an inhibitor of a disintegrin and metalloprotease 10 (ADAM10). A specific, non-limiting example of such an inhibitor is N-(3-chloro-4-methylphenyl)-2-[2-(3-methoxybenzoyl)hydrazinyl]-2-oxoacetamide (PubChem CID3117694). A similar embodiment of this method includes the further step of administering to the subject an approved anti-rheumatic drug after administering the composition or simultaneously with the composition. The approved anti-rheumatic drug can be included as a component of the composition. A preferred, non-limiting example is indomethacin.

[0010] In yet another embodiment of the method, reducing inflammation and / or treating rheumatoid arthritis includes at least one of reducing tissue swelling, lowering disease score, and decreasing the amount of at least one inflammatory biomarker. Non-limiting examples of inflammatory biomarkers are interleukin-6 (IL-6), interleukin-10 (IL-10), and C-reactive protein.

[0011] In a further aspect, the present invention provides a novel use of the disclosed pharmaceutical composition for treating immune dysfunction, autoimmunity, inflammation, and / or rheumatoid arthritis. The composition comprises a therapeutically effective dose of a modulator of a disintegrin and metalloprotease 10 (ADAM10) and at least one acceptable pharmaceutical carrier for use in a method for treating a state of immune dysfunction or inflammation (such as RA) in a subject in need of treatment. The modulator can be an inhibitor of a disintegrin and metalloprotease 10 (ADAM10). In a specific embodiment, the inhibitor of a disintegrin and metalloprotease 10 (ADAM10) is N-(3-chloro-4-methylphenyl)-2-[2-(3-methoxybenzoyl)hydrazinyl]-2-oxoacetamide (PubChem CID3117694). In another specific embodiment, the composition comprises a therapeutically effective dose of indomethacin. When used for RA, the treatment may include at least one of reducing tissue swelling, lowering RA disease score, and decreasing the amount of at least one inflammatory biomarker. The at least one inflammatory biomarker may be at least one of interleukin-6 (IL-6), interleukin-10 (IL-10), and C-reactive protein.

[0012] Other objects and advantages of this invention will become apparent from the following description, in which are set forth, by way of example, certain embodiments of the invention.

[0013] A more complete understanding of the present invention may be obtained by reference to the data set forth in the accompanying drawings, taken in conjunction with the following detailed description: Any embodiments illustrated in the drawings are intended to be merely illustrative of the present invention and should not be construed as limiting the present invention to the illustrated embodiments. [Brief description of the drawings]

[0014] [Figure 1]1 is a graph showing the disease score for CID3117694 in a rat collagen-induced arthritis model study. The disease score is used to assess the severity of rheumatoid arthritis (RA) by measuring disease activity, functional impairment, and the presence or absence of physical impairment. [Diagram 2] Graph showing paw swelling measurements for CID3117694 in a rat collagen-induced arthritis model study.* [Diagram 3] Micrographs of paw swelling observations for CID3117694 in a rat collagen-induced arthritis model study are shown. In particular, the arthritis scores of the right hind paw on day 28 of post-treatment groups (G1-G6) are shown. Three types of joints were observed for scoring: A: interphalangeal joints; B: metacarpophalangeal joints; C: carpal and tarsal joints. The following scale was used: score 0: normal; score 1: redness and swelling in one type of joint (A, B, or C); score 2: redness and swelling in two types of joints; score 3: redness and swelling in all three types of joints; score 4: maximal redness and swelling of the entire paw, losing anatomical definition. [Figure 4] Graph showing that CID3117694 dose-dependently reduces serum levels of C-reactive protein, a biomarker for RA.* [Diagram 5] Graph showing that CID3117694 dose-dependently reduces serum levels of interleukin-6 (IL-6), a biomarker for RA.* [Figure 6] 1 is a graph showing that CID3117694 dose-dependently reduces serum levels of interleukin-10 (IL-10), a biomarker for RA.* *Data shown are mean ± SD (n=6), *P<0.05, **P<0.01, ***P<0.001 indicate statistically significant, more significant, highly significant compared to disease controls (CIA Ctrl) using one-way ANOVA followed by Dunnett's test. [Figure 7] 1 shows structures of ADAM10 selective inhibitors, with the hydroxamate moiety indicated as a circle identified by an arrow. [Figure 8]100x optical micrographs of collagen-induced arthritis in the hind paws of rats from the G6(1-6) group are shown, specifically, the right hind paws of C57BL / 6 rats treated with G6[CIA+test compound 50mg / Kg]. Images shown are representative H&X stained sagittal sections of arthritic joints examined by optical microscopy. White arrows: normal healthy bone and cartilage of arthritic joints, capital letter H: hyperplasia, capital letter SI: synovial inflammation. [Figure 9] 100x optical micrographs of collagen-induced arthritis in the hind paws of rats from the G5 (1-6) group, specifically the right hind paw of a C57BL / 6 rat treated with G5 [CIA + test compound 30 mg / Kg]. Images shown are representative H&X stained sagittal sections of arthritic joints examined by optical microscopy. Capital SI: synovial inflammation, capital H: hyperplasia. [Figure 10] 100x optical micrographs of collagen-induced arthritis in the hind paws of rats from the G4(1-6) group are shown, specifically, the right hind paws of C57BL / 6 rats treated with G4[CIA+test compound 10mg / Kg]. Images shown are representative H&X stained sagittal sections of arthritic joints examined by optical microscopy. Capital SI: synovial inflammation, capital H: hyperplasia, capital P: pannus formation, capital IEJ: inflamed and eroded joints. [Figure 11] 100x optical micrographs of collagen-induced arthritis in the hind paws of rats in the G3 (1-6) group are shown, specifically, the histopathological images of the right hind paws of C57BL / 6 rats after G3 [CIA + indomethacin 2.5mg / Kg] treatment. The images shown are representative H&E stained sagittal sections of arthritic joints examined by optical microscopy. White arrows: normal morphology of bone and cartilage in arthritic joints, capital letter SI: synovial inflammation, capital letter H: hyperplasia. [Figure 12]100x light micrographs of collagen-induced arthritis in the hind paws of rats from the G2 (1-6) group are shown, specifically, the right hind paw of a C57BL / 6 rat after G2 (CIA control) treatment. Images shown are representative H&X stained sagittal sections of arthritic joints examined by light microscopy. Capital SI: synovial inflammation, capital H: hyperplasia, capital CE: cartilage erosion, capital P: pannus formation. [Figure 13] 100x optical micrographs of collagen-induced arthritis in the hind paws of rats in the G1 (1-6) group are shown, specifically, the histopathological images of the right hind paws of C57BL / 6 rats after G1 (saline) treatment. The images shown are representative H&X stained sagittal sections of arthritic joints examined by optical microscopy. White arrows: normal morphology of bone and cartilage in arthritic joints. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0015] The studies are presented and discussed herein for the purpose of promoting understanding of the principles of the present invention. Reference is made to the embodiments exemplified herein and specific language is used to describe the embodiments. However, it is understood that no limitation to the scope of the present invention is intended thereby. Any changes and further modifications in the methods, treatments, inhibitors, biomarkers, procedures, and / or compositions described, together with further applications of the principles of the present invention described herein, are expected to occur to those skilled in the art to which the present invention pertains.

[0016] A disintegrin and metaprotease 10 (ADAM10) is a member of a large group of human and non-human zinc-dependent enzymes (reviewed in Cerda-Costa and Gomis-Ruth, 2014). Structurally, it belongs to the adamalysin family (Minond, 2020, ADAM and ADAMTS enzymes). ADAM10 is a cell surface enzyme that cleaves a wide variety of cell surface proteins (Dreymueller et al., 2015; Kuhn et al., 2016; Camodeca et al., 2019; Scharfenberg et al., 2019).

[0017] ADAM10 is composed of several domains, namely, a signal sequence, a prodomain, a metalloprotease domain, a disintegrin domain, a cysteine-rich domain, a stalk region, a transmembrane domain, and a cytoplasmic tail (Minond, 2020), which are common to adamalysins (Takeda, 2009, 2016). The closest relative of ADAM10 is ADAM17, with which it shares an overall amino acid sequence identity of 24% (analysis by the Clustal Omega alignment tool). Despite the low sequence identity, ADAM10 and ADAM17 have extensively overlapping and ever-growing substrate repertoires, likely due to the lack of clear specificity of the cleavage site primary sequence (Caescu et al., 2009).

[0018] The function of ADAM10 in any particular disease or normal physiological scenario is defined by the substrates that ADAM10 cleaves. However, it is not well known whether ADAM10 and ADAM17 cleave the same substrates in the same setting. Therefore, an inhibitor selective for ADAM10 may help distinguish the role of ADAM10 in various scenarios.

[0019] The ability to cleave multiple substrates further complicates research into the role of ADAM10 and therefore its validation as a target for any particular disease. ADAM10 cleaves receptors and receptor ligands, such as cytokines, chemokines, and cell adhesion molecules, to name a few (Caescu et al., 2009; Pruessmeyer and Ludwig, 2009; Dreymueller et al., 2015; Saftig and Lichtenthaler, 2015; Moss and Minond, 2017; Wetzel et al., 2017). ADAM10-selective inhibitors that bind to the active site zinc block the proteolysis of all ADAM10 substrates. Considering that ADAM10 substrates can counteract each other's biological effects (e.g., pro- and anti-inflammatory cytokines), substrate-specific inhibitors of ADAM10 may be useful.

[0020] This finding has prompted deeper exploration of the regulatory mechanisms governing the recognition and interaction between ADAM10 and ADAM17 and their substrates. These studies have led to the realization that ADAM10 and ADAM17 may have multiple levels or ways of regulating substrate recognition and processing outside of their active sites. Among the regulatory mechanisms known so far are ADAM trafficking (Lorenzen et al., 2016; Matthews et al., 2017; Seipold et al., 2018), interactions with other proteins (Koo et al., 2020), cell membrane reorganization (Reiss and Bhakdi, 2017), non-catalytic domains of ADAMs (Willems et al., 2010; Tape et al., 2011; Stawikowska et al., 2013; Seegar et al., 2017), topology of ADAM substrates (Stawikowska et al., 2013), enzymes (Chavaroche et al., 2014), and substrate glycosylation (Minond et al., 2012). As demonstrated by several groups, these regulatory mechanisms can be targeted for modulator discovery (Tape et al., 2011; Minond et al., 2012; Madoux et al., 2016; Seegar et al., 2017).

[0021] For example, significant efforts have been dedicated to discovering modulators of ADAM10 activity for multiple indications, such as rheumatoid arthritis (RA) (Moss et al., 2008a), cancer (Moss et al., 2008b; Crawford et al., 2009; Saha et al., 2019), immune and neurodegenerative disorders (Wetzel et al., 2017). It is important to note that while some indications (e.g., Alzheimer's disease) are believed to require molecules that induce or enhance ADAM10 activity, most other indications (e.g., cancer, inflammation) require inhibitors of activity.

[0022] As shown by global knockout studies, ADAM10 (Hartmann et al., 2002) is essential for development, homeostasis, and repair, making global inhibition of all functions of this enzyme impractical as a therapeutic approach. However, tissue-specific partial knockout studies of ADAM10 (Chalaris et al., 2010) demonstrated a lack of global toxicity, suggesting that localized pharmacological partial inhibition of ADAM10 could be used.

[0023] We have discovered a new class of selective ADAM10 inhibitors (Figure 7) that act via a non-Zn-binding mechanism (Madoux et al., 2016) and potentially bind outside the active site. This non-Zn-binding mechanism of inhibition proved to be key to ensure the selectivity of these molecules over other Zn-metalloproteases. Furthermore, the lead compound of this new chemical class, CID 3117694, exhibited a unique substrate selectivity profile not observed with Zn-binding inhibitors of ADAM10 (Madoux et al., 2016), which should help avoid off-target side effects described for Zn-binding inhibitors of ADAM10. For example, inhibition of shedding of amyloid precursor protein (APP) by ADAM10 (Fahrenholz, 2007) can lead to the formation of amyloid plaques in the central nervous system. Furthermore, many of the Zn-binding inhibitors of metalloproteases caused dose-limiting toxicity known as musculoskeletal syndrome (MSS) (Overall and Lopez-Otin, 2002).

[0024] A search of the PubChem database for the biological activity of CID3117694 revealed that it was inactive in 524 bioassays and was active only against three targets by ADAM10, making it the top target (PubChem AID 743338). The second target was hERG, where CID 3117694 protected hERG from proarrhythmic drugs (PubChem AID 1511, EC50 value not reported). The third target was DNA polymerase b (PubChem AID 485314), where CID3117694 showed an IC50 value of 79 mM. It was inactive against adrenergic receptors (ADRB2), muscarinic receptors (CHRM1), and opioid receptors (OPRK1, OPRM1, and OPRD1) used in drug candidate safety screening (Bowes et al., 2012). These data suggest that CID 3117694 is a non-promiscuous compound, which should result in low off-target toxicity in vivo, and that inhibition of ADAM10 by a non-Zn-binding mechanism may be an effective strategy for treatment with fewer side effects due to superior enzyme-substrate selectivity than Zn-binding inhibitors.

[0025] Currently, there is no known RA treatment on the market or in development that targets ADAM10. Therefore, there is a need for new RA treatments. Based on this need, small molecule inhibitors of ADAM10 may be an effective preventive or therapeutic option that acts through inflammatory factors (CD23, CX3CL1 and CXCL16) that are not yet targeted by FDA-approved treatments. Inhibiting ADAM10 may have multiple benefits, including (1) reducing the levels of multiple soluble inflammatory cytokines and (2) reducing the recruitment of inflammatory cells, which is an improvement over single-target drugs (e.g., etanercept).

[0026] The main problem that seems to limit the research of ADAM10 inhibitors is the lack of target selectivity: since ADAM10 is a zinc-containing enzyme, most inhibitors bind zinc via a "warhead," and nonselective binding of such inhibitors to other zinc metalloproteases leads to intolerable side effects.

[0027] The inventors have discovered a conceptually novel class of ADAM10 inhibitors that act via a non-zinc binding inhibitory mechanism. Preliminary results obtained in different cell-based models (Table 1) demonstrated the efficacy of the lead compound CID3117694 in inhibiting inflammatory cytokines and inflammatory cell recruitment, as well as the lack of toxicity in cell-based assays.

[0028] [Table 1]

[0029] There are several selective inhibitors of ADAM10 available to researchers, including LT4 (ADAM10 IC50=40nM, ADAM17 IC50=1500nM; Zocchi et al., 2016), INCB8765 (Incyte Corporation, ADAM10 IC50=97nM, ADAM17 IC50=2045nM; Zhou et al., 2006), GI 254023X (Glaxo, ADAM10 IC50=5.3nM, ADAM17 IC50=541nM; Ludwig et al., 2005), and ADAM10 prodomain (Biozyme Inc., ADAM10 IC50=48nM, ADAM17 IC50>10mM; Moss et al., 2007). LT4, INCB8765, and GI254023X are small molecules that contain a hydroxamate moiety and therefore likely inhibit ADAM10 via a Zn-binding mechanism (Yiotakis and Dive, 2008) (Figure 7). The ADAM10 prodomain is a competitive inhibitor of ADAM10, but it is unclear whether it binds to the active site Zn. Zn-binding inhibitors may show some selectivity among closely related ADAM family members, but ultimately cannot selectively inhibit substrate shedding. There is evidence that toxicity is caused by off-target side effects resulting from the Zn-binding inhibitory mechanism that causes broad inhibition of multiple Zn metalloproteases (Dekkers et al., 1999; Newton et al., 2001; Moss and Bartsch, 2004). In addition, ADAM10 has been shown to cleave more than 70 cell surface proteins. Thus, indiscriminate inhibition of the shedding of these proteins could affect multiple biological processes (reviewed in Dreymueller et al., 2015; Wetzel et al., 2017).

[0030] A non-Zn-binding inhibitory approach for ADAM10 overcomes the shortcomings of Zn-binders. As shown by global knockout studies, ADAM10 is essential for development, homeostasis, and repair, making global inhibition of all functions of this enzyme as a therapeutic approach impractical. However, tissue-specific partial knockout studies of ADAM10 demonstrated a lack of global toxicity, suggesting that localized pharmacological partial inhibition of ADAM10 could be used. In a previous study, the inventors conducted an HTS (high-throughput screening) campaign to identify inhibitors of ADAM10 and ADAM17. This approach identified many hits with prototypic Zn-binding warheads (hydroxamic acids, polycarboxylates, polyphenols, not all shown here), which, although they showed some selectivity for ADAM10 versus ADAM17, are unlikely to have broad target selectivity. This was verified by profiling studies. However, the exceptional HTS hits are CID3117694 and related compounds (Figure 7). The exceptional HTS hits contain highly polar functionalities, but these groups are not typical Zn binders, such as the hydroxamic acid groups highlighted in LT4, INCB8765, and GI254023X. CID3117694 and related compounds are micromolar potency ADAM10 inhibitors that showed insignificant affinity for ADAM17 and a broader panel of Zn metalloproteases. This new class of selective ADAM10 inhibitors acts via a non-Zn binding mechanism, and they may bind to a previously unknown secondary substrate binding site (exosite) outside the active site.

[0031] To test the efficacy of the ADAM10 inhibitor CID3117694 for the treatment of rheumatoid arthritis (RA), the inventors performed studies using a murine collagen-induced arthritis (CIA) in vivo model. This model is the gold standard for RA preclinical testing of investigational drugs. The data obtained demonstrate that CID3117694 dose-dependently reduces RA disease score (Figure 1), paw pad swelling [Figure 2 (measurements), Figures 3A-F (observations, photographs)], and inflammatory molecules (biomarkers) (Figures 4-6). The cytokines IL-6 and IL-10 are known substrates of ADAM10, suggesting that the mechanism of action of CID3117694 is based on regulating the activity of ADAM10. Figures 8-13 are micrographs showing histopathological images of collagen-induced arthritis in the hind paws of the test groups of rats.

[0032] conclusion A disintegrin and metalloprotease 10 (ADAM10) is an important target for several therapeutic agents, especially anti-rheumatic drugs. Overall, the present study shows that the ADAM10 selective inhibitor N-(3-chloro-4-methylphenyl)-2-[2-(3-methoxybenzoyl)hydrazinyl]-2-oxoacetamide (PubChem CID3117694) overcomes the shortcomings of previous anti-rheumatic compounds and can provide a novel treatment for rheumatoid arthritis with increased efficacy.

[0033] All patents and publications mentioned in this specification are indicative of the level of skill of those skilled in the art to which the invention pertains. All patents and publications are incorporated herein by reference to the same extent as if each individual publication was specifically and individually indicated to be incorporated by reference. It should be understood that, while specific forms of the invention have been illustrated, they are not intended to be limited to the specific forms or arrangements described and illustrated herein. It will be apparent to those skilled in the art that various modifications may be made without departing from the scope of the invention, and the invention should not be considered as limited to those shown and described in the specification. Those skilled in the art will readily appreciate that the invention is well adapted to carry out the objects and obtain the objects and advantages mentioned, as well as those inherent therein. The methods, treatments, inhibitors, biomarkers, procedures, and / or compositions described herein are presently representative of preferred embodiments and are intended to be exemplary, and not limiting in scope. Modifications therein and other uses encompassed within the spirit of the invention will occur to those skilled in the art. Although the invention has been described in connection with specific preferred embodiments, it is understood that the invention as ultimately claimed should not be unduly limited to such specific embodiments. Indeed, various modifications of the described modes for carrying out the invention which are obvious to those skilled in the art are intended to be within the scope of the present invention.

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[0035] STATEMENT REGARDING FEDERALLY SPONSORED RESEARCH OR DEVELOPMENT This invention was made with government support under Grant No. DA033985 awarded by the National Institutes of Health. The United States Government has certain rights in the invention.

Claims

1. 1. A method for treating a condition of immune dysfunction in a subject in need thereof, comprising: providing a composition comprising a therapeutically effective dose of a modulator of a disintegrin and metalloprotease 10 (ADAM10) and at least one acceptable pharmaceutical carrier; administering the composition to the subject to treat the condition of immune dysfunction in the subject through modulation of ADAM10; The method includes:

2. The method of claim 1 , wherein the modulator is an inhibitor of a disintegrin and metalloprotease 10 (ADAM10).

3. 3. The method of claim 2, wherein said inhibitor of a disintegrin and metalloprotease 10 (ADAM10) is N-(3-chloro-4-methylphenyl)-2-[2-(3-methoxybenzoyl)hydrazinyl]-2-oxoacetamide (PubChem CID 3117694).

4. 3. The method of claim 2, wherein said condition of immune dysfunction is rheumatoid arthritis (RA).

5. 5. The method of claim 4, further comprising administering to the subject a therapeutically effective dose of indomethacin.

6. 6. The method of claim 5, wherein the therapeutically effective dose of indomethacin is contained in the composition.

7. 1. A method for reducing inflammation in a subject in need of treatment, comprising: providing a composition comprising a therapeutically effective dose of a modulator of a disintegrin and metalloprotease 10 (ADAM10) and at least one acceptable pharmaceutical carrier; administering the composition to the subject to reduce inflammation in the subject through modulation of ADAM10; The method includes:

8. The method of claim 7, wherein the modulator is an inhibitor of a disintegrin and metalloprotease 10 (ADAM10).

9. 9. The method of claim 8, wherein said inhibitor of a disintegrin and metalloprotease 10 (ADAM10) is N-(3-chloro-4-methylphenyl)-2-[2-(3-methoxybenzoyl)hydrazinyl]-2-oxoacetamide (PubChem CID 3117694).

10. 9. The method of claim 8, wherein the inflammation is a result of rheumatoid arthritis.

11. 11. The method of claim 10, wherein the reduction in inflammation comprises at least one of a reduction in tissue swelling, a lowered disease score, and a reduced amount of at least one inflammatory biomarker.

12. 12. The method of claim 11, wherein the at least one inflammatory biomarker is at least one of interleukin-6 (IL-6), interleukin-10 (IL-10), and C-reactive protein.

13. 10. The method of claim 8, further comprising administering to the subject a therapeutically effective dose of indomethacin.

14. 14. The method of claim 13, wherein the therapeutically effective dose of indomethacin is contained in the composition.

15. 1. A method for treating rheumatoid arthritis in a subject in need thereof, comprising: providing a composition comprising a therapeutically effective dose of a modulator of a disintegrin and metalloprotease 10 (ADAM10) and at least one acceptable pharmaceutical carrier; administering the composition to the subject to treat rheumatoid arthritis in the subject through modulation of ADAM10; The method includes:

16. 16. The method of claim 15, wherein the modulator is an inhibitor of a disintegrin and metalloprotease 10 (ADAM10).

17. 17. The method of claim 16, wherein said inhibitor of a disintegrin and metalloprotease 10 (ADAM10) is N-(3-chloro-4-methylphenyl)-2-[2-(3-methoxybenzoyl)hydrazinyl]-2-oxoacetamide (PubChem CID 3117694).

18. 17. The method of claim 16, wherein the treatment of rheumatoid arthritis (RA) comprises at least one of reducing tissue swelling, lowering RA disease score, and decreasing the amount of at least one inflammatory biomarker.

19. 19. The method of claim 18, wherein the at least one inflammatory biomarker is at least one of interleukin-6 (IL-6), interleukin-10 (IL-10), and C-reactive protein.

20. 17. The method of claim 16, further comprising administering to the subject a therapeutically effective dose of indomethacin.

21. 21. The method of claim 20, wherein the therapeutically effective dose of indomethacin is contained in the composition.

22. 1. A method for treating rheumatoid arthritis in a subject in need thereof, comprising: providing a composition comprising a therapeutically effective dose of an inhibitor of a disintegrin and metalloprotease 10 (ADAM10) and at least one acceptable pharmaceutical carrier; administering the composition to the subject to treat rheumatoid arthritis in the subject by inhibiting ADAM10; The method includes:

23. 23. The method of claim 22, wherein said inhibitor of a disintegrin and metalloprotease 10 (ADAM10) is N-(3-chloro-4-methylphenyl)-2-[2-(3-methoxybenzoyl)hydrazinyl]-2-oxoacetamide (PubChem CID 3117694).

24. 24. The method of claim 23, wherein the treatment of rheumatoid arthritis (RA) comprises at least one of reducing tissue swelling, lowering RA disease score, and reducing the amount of at least one inflammatory biomarker.

25. 25. The method of claim 24, wherein the at least one inflammatory biomarker is at least one of interleukin-6 (IL-6), interleukin-10 (IL-10), and C-reactive protein.

26. 24. The method of claim 23, further comprising administering to the subject a therapeutically effective dose of indomethacin.

27. 27. The method of claim 26, wherein the therapeutically effective dose of indomethacin is contained in the composition.

28. 1. A method for treating rheumatoid arthritis in a subject in need thereof, comprising: providing a composition comprising a therapeutically effective dose of N-(3-chloro-4-methylphenyl)-2-[2-(3-methoxybenzoyl)hydrazinyl]-2-oxoacetamide (PubChem CID 3117694) and at least one acceptable pharmaceutical carrier; administering the composition to the subject to treat rheumatoid arthritis in the subject; The method includes:

29. 30. The method of claim 28, wherein the treatment of rheumatoid arthritis (RA) comprises at least one of reducing tissue swelling, lowering RA disease score, and reducing the amount of at least one inflammatory biomarker.

30. 30. The method of claim 29, wherein the at least one inflammatory biomarker is at least one of interleukin-6 (IL-6), interleukin-10 (IL-10), and C-reactive protein.

31. 30. The method of claim 28, further comprising administering to the subject a therapeutically effective dose of indomethacin.

32. 32. The method of claim 31 , wherein the therapeutically effective dose of indomethacin is contained in the composition.

33. A composition comprising a therapeutically effective dose of a modulator of a disintegrin and metalloprotease 10 (ADAM10) and at least one acceptable pharmaceutical carrier for use in a method for treating a condition of immune dysfunction or inflammation in a subject in need of such treatment.

34. 34. The use according to claim 33, wherein the modulator is an inhibitor of a disintegrin and metalloprotease 10 (ADAM10).

35. 35. The use according to claim 34, wherein said inhibitor of a disintegrin and metalloprotease 10 (ADAM10) is N-(3-chloro-4-methylphenyl)-2-[2-(3-methoxybenzoyl)hydrazinyl]-2-oxoacetamide (PubChem CID 3117694).

36. 35. The use according to claim 34, wherein said condition of immune dysfunction or inflammation is rheumatoid arthritis (RA).

37. A composition comprising a therapeutically effective dose of a modulator of a disintegrin and metalloprotease 10 (ADAM10), a therapeutically effective dose of indomethacin, and at least one acceptable pharmaceutical carrier for use in a method for treating a condition of immune dysfunction or inflammation in a subject in need of treatment.

38. 38. The use according to claim 37, wherein the modulator is an inhibitor of a disintegrin and metalloprotease 10 (ADAM10).

39. 39. The use according to claim 38, wherein said inhibitor of a disintegrin and metalloprotease 10 (ADAM10) is N-(3-chloro-4-methylphenyl)-2-[2-(3-methoxybenzoyl)hydrazinyl]-2-oxoacetamide (PubChem CID 3117694).

40. 38. The use according to claim 37, wherein said condition of immune dysfunction or inflammation is rheumatoid arthritis (RA).

41. A composition comprising a therapeutically effective dose of N-(3-chloro-4-methylphenyl)-2-[2-(3-methoxybenzoyl)hydrazinyl]-2-oxoacetamide (PubChem CID 3117694) and at least one acceptable pharmaceutical carrier for use in a method for treating rheumatoid arthritis in a subject in need of such treatment.

42. 42. The use of claim 41, wherein the treatment of rheumatoid arthritis (RA) comprises at least one of reducing tissue swelling, lowering RA disease score, and reducing the amount of at least one inflammatory biomarker.

43. 43. The use of claim 42, wherein the at least one inflammatory biomarker is at least one of interleukin-6 (IL-6), interleukin-10 (IL-10) and C-reactive protein.

44. 1. A composition comprising a therapeutically effective amount of N-(3-chloro-4-methylphenyl)-2-[2-(3-methoxybenzoyl)hydrazinyl]-2-oxoacetamide (PubChem CID 3117694), a therapeutically effective dose of indomethacin, and at least one acceptable pharmaceutical carrier for use in a method for treating rheumatoid arthritis in a subject in need of such treatment.

45. 45. The use of claim 44, wherein the treatment of rheumatoid arthritis (RA) comprises at least one of reducing tissue swelling, lowering RA disease score, and reducing the amount of at least one inflammatory biomarker.

46. 46. ​​The use of claim 45, wherein the at least one inflammatory biomarker is at least one of interleukin-6 (IL-6), interleukin-10 (IL-10) and C-reactive protein.

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