Description:FORM 2
The Patents Act 1970
(39 of 1970)
&
The Patent Rules 3003
COMPLETE SPECIFICATION
(See Section 10 and rule 13)
1. TITLE: THERAPEUTIC TARGET FOR TREATMENT OF RHEUMATOID ARTHRITIS (RA)
2. APPLICANT (s):
Name in Full Nationality Address of the Applicant
UNIVERSITY OF DELHI INDIA House No. Department Of Genetics, University of Delhi, South Campus, Delhi- 110021, India
Street
City
State
Country
Pin code
3. PREAMBLE OF THE DESCRIPTION: The following COMPLETE specification particularly describes the disclosure and the manner in which it is performed.
FIELD OF THE DISCLOSURE
The present disclosure generally relates to treatment of Rheumatoid Arthritis (RA). In particular, the present disclosure relates a therapeutic target for treatment of RA
BACKGROUND
Rheumatoid Arthritis (RA) is an autoimmune disease, characterized by a chronic inflammation of various organs, mainly joints along with immune dysregulation. RA synovial fibroblasts are one of the major cell types involved in bone and cartilage destruction. Cytokines, including TNF alpha and IL-6, are involved in the pathogenesis and maintenance of the disease. There is no cure for RA till date.
There are many compositions available in the art to treat RA. However, such medications actually do not treat RA and tend to reduce effects of RA in joints. For example, reducing pain and stiffness in affected joints, preventing joint damage as much as possible, minimizing any disability caused by pain, joint damage, or deformity and reducing the risk of developing associated conditions such as heart disease. Medicines which are advised to ease pain and stiffness include non-steroidal anti-inflammatory drugs (NSAIDs). Role thereof is limited as such drugs do not have any effect on disease progression.
Second line of drugs include 'disease-modifying anti-rheumatic drugs' (DMARDs, such as methotrexate. Such drugs ease symptoms and work by reducing joint inflammation. There are many non-responders to these drugs making it a difficult choice for clinicians.
Next set of drugs called biological DMARDs have recently been developed and they include anti-TNF, IL6 blockers like etanercept, infliximab etc. Though such drugs show promise but long-term benefits thereof are still being evaluated as well as they are very expensive. Use of such drugs against cytokines such as TNF alpha have been shown to be associated with the risk of infection and malignancies.
Along with some serious side effects of the said medications, medication withdrawal after the remission has also not been proven beneficial for the patients, making RA a life-long condition that requires constant and expensive management. Therefore, search for causative agents and better therapeutics based on biologics like antibodies need to be done for developing a potential treatment for RA.
Therefore, there exists a need for developing alternatives to get rid of aforementioned issues.
OBJECTS OF THE EMBODIMENT
One object of the present disclosure is to disclose a therapeutic target for treatment of Rheumatoid Arthritis (RA).
Another object of the present disclosure is to disclose the therapeutic target for treatment of Rheumatoid Arthritis (RA) that includes a G-protein ARL15 downregulated.
Another object of the present disclosure is to disclose the G-protein ARL15 downregulated to reduce redness and inflammation in RA.
Another object of the present disclosure is to disclose the G-protein ARL15 downregulated does not follow TNF pathway.
Another object of the present disclosure is to disclose monoclonal antibodies target G-protein ARL15.
Another object of the present disclosure is to disclose the ARL15mAb is non-toxic to subject.
Another object of the present disclosure is to disclose the ARL15mAb decreases synovitis formation thereby decreasing the flux of invading synovial fibroblasts.
Another object of the present disclosure is to disclose the ARL15mAb minimizes bone degradation in RA.
In this respect, before explaining at least one embodiment of the present disclosure in detail, it is to be understood that the disclosure is not limited to in its application to the details of processing and to the arrangements of the components set forth in the following description or illustrated in the drawings. The disclosure is capable of embodiments in addition to those described and of being practised and carried out in various ways. Also, it is to be understood that the phraseology terminology employed herein, as well as the abstract, are for the purpose of description and should not be regarded as limiting.
SUMMARY
In an embodiment, the present disclosure discloses a therapeutic target for treatment of Rheumatoid Arthritis (RA). The target includes a G-protein ARL15 downregulated to reduce redness and inflammation in RA. A monoclonal antibody targets ARL15 to obtain ARL15mAb. The ARL15mAb does not follow TNF pathway. The ARL15mAb negates side effects of cytokine blockage. The ARL15mAb is non-toxic to animals.
BRIEF DESCRIPTION OF THE DRAWINGS
Other objects, features, and advantages of the embodiment will be apparent from the following description when read with reference to the accompanying drawings. In the drawings, wherein like reference numerals denote corresponding parts throughout the several views:
Referring to Figure 1, shows an effect of mAb1 on Clinical score on Day 14; Data is represented as Mean ± SEM (n=3-12), *p<0.05, vs Isotope control, IP; $ p<0.05, vs Isotope control, IV; ANOVA followed by Dunnett's test, in accordance with an illustrative embodiment of a present disclosure;
Referring to Figure 2, shows an effect of anti-mouse type II collagen IgG1 antibody; Data is represented as Mean ± SEM (n=3-12), *p<0.05, vs Isotope control, IP; $ p<0.05, vs Isotope control, IV; ANOVA followed by Dunnett's test, in accordance with the illustrative embodiment of the present disclosure;
Referring to Figure 3, shows an effect of mAb1 on Serum Pro-inflammatory Cytokines IL-17; Data is represented as Mean ± SEM (n=3-12), *p<0.05, vs Isotope control, IP; ANOVA followed by Dunnett’s multiple comparison test, in accordance with the illustrative embodiment of the present disclosure;
Referring to Figure 4, shows an effect of mAb1 on Serum Pro-inflammatory Cytokines IL-6; Data is represented as Mean ± SEM (n=3-12), *p<0.05, vs Isotope control, IP; ANOVA followed by Dunnett’s multiple comparison test, in accordance with the illustrative embodiment of the present disclosure; and
Referring to Figure 5, shows an effect of mAb1 on Serum Pro-inflammatory Cytokines TNF-a; Data is represented as Mean ± SEM (n=3-12), *p<0.05, vs Isotope control, IP; ANOVA followed by Dunnett’s multiple comparison test, in accordance with the illustrative embodiment of the present disclosure.
Sequences
Seq 1: >NM_019087.3 Homo sapiens ADP ribosylation factor like GTPase 15 (ARL15), mRNA; and
Seq 2: >NP_061960.1 ARL15 [organism=Homo sapiens] [GeneID=54622]
DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS
The embodiments herein and the various features and advantageous details thereof are explained more fully with reference to the non-limiting embodiments that are illustrated in the accompanying drawings and detailed in the following description. Descriptions of well-known components and processing techniques are omitted so as to not unnecessarily obscure the embodiments herein. The examples used herein are intended merely to facilitate an understanding of ways in which the embodiments herein may be practiced and to further enable those of skill in the art to practice the embodiments herein. Accordingly, the examples should not be construed as limiting the scope of the embodiments herein.
Many modifications will be apparent to those skilled in the art without departing from the scope of the present invention as hereinbefore described with reference to the accompanying drawings.
Throughout this specification and the claims which follow, unless the context requires otherwise, the word “comprise”, and variations such as “comprises” and “comprising”, will be understood to imply the inclusion of a stated integer or step or group of integers or steps but not the exclusion of any other integer or step or group of integers or steps.
As used herein, the singular forms “a”, “an”, “the” include plural referents unless the context clearly dictates otherwise. Further, the terms “like”, “as such”, “for example”, “including” are meant to introduce examples which further clarify more general subject matter, and should be contemplated for the persons skilled in the art to understand the subject matter.
It is to be noted that the Seq 1 and Seq 2 are well known sequences already deposited in the NCBI database https://www.ncbi.nlm.nih.gov/datasets/gene/id/54622/products/.
The present disclosure discloses a therapeutic target for treatment of Rheumatoid Arthritis (RA). The target includes a G-protein ARL15 downregulated to reduce redness and inflammation in RA. The ARL15 is a non-HLA gene, overexpressed in RA. Knockdown of such gene in patient derived cell lines down regulates the expression of genes which are critical for RA.
A monoclonal antibody targets ARL15 to obtain ARL15mAb. The ARL15mAb does not follow TNF pathway. The ARL15mAb negates side effects of cytokine blockage. The ARL15mAb is non-toxic to animals.
Example1
DBA1/J female mice obtained for the study were randomly divided into four groups and each of them was marked by ear punch. The groups were termed healthy, vehicle control, ARL15 mAb and Isotype control. On day 1 after acclimatization of 7 days, all DBA/1 mice except those in the healthy group were immunized by intradermal injections of an emulsion containing 100 µg of immunization grade bovine type II collagen in complete Freund's adjuvant containing heat-killed Mycobacterium butyricum the base of the tail. On day 21, a booster containing 100 µg bovine CII in incomplete Freund's adjuvant was administered following previously described protocol (ref). The animals were being observed for disease development. Treatment was initiated after 12 days of the booster dose when disease severity of all groups was observed to be maximum. Drugs were administered intraperitoneally (IP) three times in a week. The healthy group did not receive any treatment. To assess the disease characteristics, all the mice were scored for disease extent at 4-5 days intervals for ~20 days following the booster immunization. both paws and joints of fore and hind limbs were scored as follows: 0- Normal; 1- swelling and redness of the paw or one digit; 2- swelling and redness of ankle and wrist; 3- severe redness and swelling of entire paw including digits and 4- severe arthritis of the limb including entire paw and digits.
Example2
The levels of IL6 and TNFa in the sera samples of the experimental animals were measured using mouse TNFa and IL6 ELISA kits (BMS607HS & KMC0061, Thermo, USA). Briefly, standards in 500-7.8 pg/ml range were prepared using serial dilution of a 1000pg vial. The sera samples were diluted as per manufacturer’s instruction and 100µl of samples along with the diluted standards were loaded in the well of ELISA strips. Biotin conjugate was then added to each of the wells followed by incubation for 2 hours at room temperature. Color was developed in the wells by treatment of streptavidin-HRP followed by TMB substrate solution. OD readings in each of the wells were captured at 450nm on spectrophotometer and the concentrations of cytokines were derived based on the values of standards.
Figure 1 shows lower level of IgG1 on treatment with mAb1. Figure 2 shows significantly lower level of IL17 on treatment with mAb1. Figure 3 shows very low level of IL6 on treatment with mAb1. Figure 4 shows lower level of IL1 on treatment with mAb1. Figure 5 shows significant reduction of TNF level on treatment with mAb1.
The foregoing descriptions of exemplary embodiments of the present disclosure have been presented for purposes of illustration and description. They are not intended to be exhaustive or to limit the disclosure to the precise forms disclosed, and obviously many modifications and variations are possible in light of the above teaching. The exemplary embodiments were chosen and described in order to best explain the principles of the disclosure and its practical application, to thereby enable others skilled in the art to best utilize the disclosure and various embodiments with various modifications as are suited to the particular use contemplated. It is understood that various omissions, substitutions of equivalents are contemplated as circumstance may suggest or render expedient but is intended to cover the application or implementation without departing from the spirit or scope of the claims of the present disclosure.
, Claims:We Claim
1. A therapeutic target for treatment of Rheumatoid Arthritis (RA), the target comprising a G-protein ARL15 downregulated to reduce redness and inflammation in RA.
2. The target as claimed in claim 1, wherein a monoclonal antibody targets ARL15 to obtain ARL15mAb.
3. The target as claimed in claim 2, wherein the ARL15mAb does not follow TNF pathway.
4. The target as claimed in claim 2, wherein the ARL15mAb negates side effects of cytokine blockage.
5. The target as claimed in claim 2, wherein the ARL15mAb is non-toxic to animals.