Patent Application
20220323481
The present invention relates to a pharmaceutical composition comprising as active ingredients thereof siRNA and a DNA vector capable of expressing that siRNA, and more particularly, relates to inhibition of expression of human CHST15, a pharmaceutical composition comprising as active ingredients thereof siRNA and a DNA vector capable of expressing that siRNA, the use thereof, and an administration method.
Many chronic diseases are autoimmune diseases that ae associated with chronic inflammation. In recent years, attention has focused on the invasion of tissue by leukocytes involved in inflammation as a novel treatment target for these chronic diseases. The invasion of inflammatory tissue by leukocytes circulating in the blood consists of the four stages indicated below. Namely, (1) decrease in leukocyte flow rate due to a first interaction between vascular endothelial cells and leukocytes in the vicinity of the inflammation site (rolling), (2) activation of rolling leukocytes, (3) strong adhesion of the aforementioned activated leukocytes to the aforementioned vascular endothelial cells due to a second interaction in the vicinity of the inflammation site, and (4) final migratory invasion of the aforementioned activated leukocytes into tissue by slipping through blood vessels by passing between vascular endothelial cells. Among these stages, bonding between L-selectin on the surface of the leukocytes and the end of a 6-sulfosiallyl Lewis X-type sugar chain of L-selectin ligand present on the surface of vascular endothelial cells is known to be involved in the first stage rolling. N-acetylglucosamine-6-sulfotransferase is known to be involved as the enzyme involved in inflammation site-specific synthesis of this 6-sulfosiallyl Lewis X-type sugar chain, and the sulfotransferase encoded by human CHST2 and CHST4 genes has conventionally been thought to be involved in invasion of inflammatory tissue by leukocytes circulating in the blood. However, based on research conducted on knockout mice, although circulating leukocytes invade inflammatory tissue even in mice lacking both CHST2 and CHST4 genes, the reason for this has not been clearly determined (Non-Patent Document 1).
The inventors of the present invention reported therapeutic effects, including inhibition of ulceration, inflammation and fibrosis, using siRNA expressing sulfotransferase CHST15 that is different from CHST2 and CHST4 (Patent Documents 1 to 3 and Non-Patent Documents 1 and 2). In particular, a phase IIa clinical trial was recently conducted on human Crohn's disease patients, and it was demonstrated that when the aforementioned siRNA was administered by submucosal administration into the large intestine of these patients, healing of the mucosa or healing of ulceration was able to be achieved endoscopically.
During the course thereof, since siRNA therapy using CHST15 yields superior treatment results in comparison with conventional biological preparation therapy for Crohn's disease, it was found that siRNA of CHST15 inhibits the first stage of invasion of inflammatory tissue by circulating leukocytes, thereby leading to completion of the present invention.
It is necessary to develop a novel treatment method for chronic disease for which conventional treatment methods are either ineffective or for which efficacy is low.
Since siRNA that inhibits expression of CHST15 inhibits expression of 6-sulfosiallyl Lewis X of the L-selectin ligand of vascular endothelial cells at an inflammation site, a novel technology for chronic disease was developed using the aforementioned siRNA that can be used in combination with conventional therapy.
The present invention provides a pharmaceutical composition for the treatment and/or prevention of an inflammatory chronic disease that is used in combination with a biological preparation inhibiting leukocyte tissue invasion and/or a biological preparation inhibiting inflammatory cytokines. The pharmaceutical composition of the present invention contains as active ingredients thereof:
(i) siRNA suppressing the expression of CHST15 gene that contains a structure formed by the hybridization of RNA containing the base sequence represented by SEQ ID NO: 1 with RNA containing the base sequence complementary thereto,
(ii) the siRNA of (i) having a structure in which one or a plurality of nucleic acids overhang from the end thereof, or
(iii) a DNA vector capable of expressing the siRNA of (i) or (ii).
In the pharmaceutical composition of the present invention, the biological preparation inhibiting leukocyte tissue invasion may inhibit the function of at least one molecule selected from the group consisting of integrin and/or chemokine receptor on the surface of leukocytes circulating in the blood and adhesion molecules on the surface of vascular endothelial cells.
In the pharmaceutical composition of the present invention, the biological preparation inhibiting leukocyte tissue invasion may be at least one member selected from the group consisting of Etrolizumab, Vedolizumab, Natalizumab, PF-00547659 and Vercirnon.
In the pharmaceutical composition of the present invention, the biological preparation inhibiting inflammatory cytokines may inhibit the function of at least one molecule selected from the group consisting of TNF-α, IL-17 and IL-23.
The pharmaceutical composition of the present invention may further use in combination at least one member selected from the group consisting of a 5-aminosalicyclic acid preparation, steroid preparation, thiopurine preparation, and immunosuppressants including tacrolimus.
The present invention provides a pharmaceutical composition for the treatment and/or prevention of chronic disease in which 6-sulfosiallyl Lewis X of L-selectin ligand is expressed on the surface of the vascular endothelial cells of a patient. The pharmaceutical composition of the present invention contains as active ingredients thereof:
(i) siRNA suppressing the expression of CHST15 gene that contains a structure formed by the hybridization of RNA containing the base sequence represented by SEQ ID NO: 1 with RNA containing the base sequence complementary thereto,
(ii) the siRNA of (i) having a structure in which one or a plurality of nucleic acids overhang from the end thereof, or
(iii) a DNA vector capable of expressing the siRNA of (i) or (ii).
In the pharmaceutical composition of the present invention, the chronic disease may be an autoimmune disease.
In the pharmaceutical composition of the present invention, the autoimmune disease may be at least one disease selected from the group consisting of inflammatory colitis, Crohn's disease, ulcerative colitis, autoimmune pancreatitis, chronic rheumatoid arthritis, bronchial asthma, chronic interstitial pneumonia, Grave's disease, Hashimoto's thyroiditis, chronic thyroiditis and atopic dermatitis.
The pharmaceutical composition of the present invention may be administered systemically or locally.
In the pharmaceutical composition of the present invention, the autoimmune disease may be selected from the group consisting of inflammatory colitis, Crohn's disease and ulcerative colitis, and the local administration may be submucosal administration into the intestine of a patient.
In the pharmaceutical composition of the present invention, the systemic administration may be oral administration and/or intravenous injection.
In the pharmaceutical composition of the present invention, a complex may be administered orally that contains N-acetylated chitosan and an active ingredient in the form of:
(i) siRNA suppressing expression of CHST15 gene that contains a structure obtained as a result of hybridization of RNA containing the base sequence represented by SEQ ID NO: 1 with RNA containing the base sequence complementary thereto, or
(ii) the siRNA of (i) having a structure in which one or a plurality of nucleic acids overhang from the end thereof.
All publications mentioned in the present description are incorporated in the present description in their entirety by reference.
In the present invention, “administering in combination” refers to the administration of the pharmaceutical composition of the present invention which is administered simultaneously, continuously or after allowing an certain period of time after having administration of the other, a biological preparation that inhibits tissue invasion by leukocytes and/or a biological preparation that inhibits inflammatory cytokines, optionally 5-aminosalicylic acid, steroid preparations, thiopurine preparations and immunosuppressants. In the case of administering a biological preparation that inhibits tissue invasion by leukocytes and/or a biological preparation that inhibits inflammatory cytokines, and optionally at least one member selected from the group consisting of 5-aminosalicylic acid, steroid preparations, thiopurine preparations and immunosuppressants, and the pharmaceutical composition of the present invention, overlapping of the administration period of the pharmaceutical composition of the present invention and the administration period of either of the aforementioned biological preparations, or administration of the pharmaceutical composition of the present invention within a period equal to at least 20% of the administration period of the biological preparation following completion of the administration period of the biological preparation, is included in “administration in combination”. Although the dose of the pharmaceutical composition of the present invention can be suitably adjusted according to such factors as body weight, age and symptoms of the subject to receive administration, in the case, for example, the biological preparation is an antibody, the dose is, for example, 0.1 mg/kg/week to 100 mg/kg/week or a dose that yields a blood concentration equivalent thereto, preferably 1 mg/kg/week to 50 mg/kg/week or a dose that yields a blood concentration equivalent thereto, and more preferably 5 mg/kg/week to 10 mg/kg/week or a dose that yields a blood concentration equivalent thereto. In addition, when the antimetabolite is gemcitabine hydrochloride, the dose is, for example, 10 mg/m2/week to 10000 mg/m2/week or a dose that yields a blood concentration equivalent thereto, preferably 100 mg/m2/week to 5000 mg/m2/week or a dose that yields a blood concentration equivalent thereto, and more preferably 500 mg/m2/week to 1500 mg/m2/week or a dose that yields a blood concentration equivalent thereto.
An administration method, administration interval and dose that yield a therapeutic effect similar to the effect of the present invention can be suitably selected for the aforementioned administration method, administration interval and dose. For example, an administration method, administration interval and dose that yield an effect similar to that of the aforementioned preferable examples can be selected by measuring the blood concentrations of a biological preparation that inhibits tissue invasion by leukocytes and/or a biological preparation that inhibits inflammatory cytokines, at least one member selected from the group consisting of 5-aminosalicylic acid, steroid preparations, thiopurine preparations and immunosuppressants depending on the case, and the pharmaceutical composition of the present invention, and an administration method, administration interval and dose that achieve a blood concentration equivalent to that of the aforementioned examples are included in the present invention.
In the present description, examples of diseases used for treatment and/or prevention by the pharmaceutical composition of the present invention include, but are not limited to, Guillain-Barré syndrome, myasthenia gravis, chronic gastritis, chronic atrophic gastritis, autoimmune hepatitis, primary biliary cholangitis, inflammatory bowel disease, Crohn's disease, ulcerative colitis, autoimmune pancreatitis, Takayasu's arteritis, Goodpasture's syndrome, rapidly progressive glomerulonephritis, Grave's disease, Hashimoto's thyroiditis, primary hypothyroidism, idiopathic Addison's disease, type 1 diabetes, chronic discoid lupus erythematosus, localized scleroderma, pemphigus, pustular psoriasis, psoriasis vulgaris, acquired epidermolysis bullosa, autoimmune optic neuropathy, chronic rheumatoid arthritis, systemic lupus erythematosus, scleroderma, polymyositis, dermatomyositis, vasculitis syndrome, mixed connective tissue disease, bronchial asthma, chronic thyroiditis and atopic dermatitis.
A detailed explanation of the present invention and its examples are also explained by the following references included in the submitted documents of the present application and documents cited therein.
In particular, references consisting of Suzawa, K., et al., Am. J. Gastroenterol. 2007; 102: 1499-1509, Yeh, J. C., et al., Cell, 2001; 105: 957-969, Kobayashi, M., et al., Biol. Pharm. Bull. 2009; 32: 774-779, and A. van Zante and S. D. Rosen, Biochemical Society Transactions 2003; 31: 313-317 are incorporated in their entirety in the present description by reference.
8.1 Study Design
This is a randomized, double-blinded, placebo-controlled, parallel group study or STNM01 by a single dose submucosal injection.
Freeze-dried STNM01 will be diluted with physiological saline. Study drugs will be administered submucosally using an endoscope to the rectosigmioidal region. The patients will be randomized to receive either STNM01 (in concentrations of 25 nM and 250 nM) or placebo. Eligible subjects will be admitted to the study sites and receive a single dose of the study drug on Day 1. Subjects will be discharged alter confirming that there are no safety concerns on Day 2. They will return to the study sites for follow-up examinations 14 and 28 days after administration.
Table 8-1 shows the dose (concentration), dosage regimen and number of subjects.
Local administration of STNM01 (including 25 nM and 250 nM) in the completed randomized, double-blind, placebo-controlled Phase 1 clinical study was generally well tolerated. Therefore, there is no safety concern that would prevent the conduct of the proposed, parallel group Phase 2a clinical study at this point of time. Mucosal healing efficacy observed in the Phase 1 study was unexpected as the primary objective of the Phase 1 study was to evaluate safety. Thus, the planned Phase 2a study is intended to confirm the efficacy observed in the Phase 1 study and to gain more information about efficacy parameters and the doses to be used in a larger controlled next study to be performed. Although this is a parallel group-comparison design, safety of the patients will be closely monitored and will be assessed as a secondary endpoint.
8.2 Discontinuation or Interruption of the Study
When one or more subjects in the current step experienced a serious or a severe adverse event which is considered to have been caused by factors other than the study drug, e.g., a study procedure, or, when half or more of the subjects in the current step experienced a moderate adverse event requiring medical intervention which is considered to have been caused by factors other than the study drug, the sponsor will temporarily discontinue the study and review the study continuation in reference to the principal investigator's opinion.
8.3 Planned Sample Size
A two group Chi-Square test with a 0.050 two-sided significance level will have 80% power to detect the difference between active treatment and placebo (89% versus 11%) when the sample size in each group is 8 (assuming a withdrawal rate of 15%).
Therefore, eight subjects per group (8 for STNM01 25 nM, 8 for STNM01 250 nM, 8 for placebo), 24 subjects in total will be included.
8.4 Planned Duration of Study
May 2013 to May 2016
8.5 Discussion of Study Design and Choice of Control Groups
This is a P2a study to investigate the safety and efficacy of a single dose of STNM01. The design of the study is deemed appropriate for this type of study.
Participation in the study will be offered to all patients who are eligible according to the inclusion and exclusion criteria, who are patients of or referred to the participating institutions and who are presented to the study investigators.
9.1 Indication
Patients with Ulcerative Colitis with endoscopic active lesion(s).
9.2 Inclusion and Exclusion Criteria
9.2.1 Inclusion Criteria
Subject eligibility is determined according to the following criteria.
9.2.2. Exclusion Criteria
Any subject who meets any of the following criteria will not qualify for entry into the study:
9.3 Prior and Concomitant Medications and Therapies
Concomitant use of prior medications and therapies with STNM01 in this study can be justified because there were very few toxic findings and the drug eliminated from blood rapidly after administration in the non-clinical studies of STNM01, thus we speculate that onset of unknown adverse drug reactions or increase in severity of adverse drug reactions of the prior medications will be unlikely.
9.4 Restrictions During the Study
Subjects must be instructed to follow the instructions below. They will stay under medical supervision of the investigator during hospitalization.
9.4.1 Food, Drinks, Smoking and Exercise
Subjects will start fasting after evening meal on Day 0 through 4 hours after study drug administration on Day 1. They will also start fasting after evening meal before the day of follow-up examinations until their completion.
Food and drinks containing alcohol and caffeine will be prohibited during hospitalization. After discharge, excessive drinking will be prohibited until the end of final examinations/observations. Alcohol beverages will not be allowed from the day before follow-up examinations until their completion.
Food and beverages other than those to be provided in the study site will not be allowed during hospitalization. Subjects must be instructed to refrain horn excessive earing and drinking after discharge until the end of final examinations/observations.
Smoking will be prohibited during hospitalization. Smoking will not be allowed from bedtime on the day before follow-up examinations until their completion.
Physical exercise will be prohibited while subjects are in the study site as well as from the day before follow-up examinations until their completion.
9.4.2 Contacts to Subjects after Discharge
The site management organization will contact the subjects in a timely manner prior to the scheduled visit regarding the study procedure they need to follow (e.g. restrictions on excessive eating and drinking after discharge).
9.5 Criteria for Discontinuation or Withdrawal of a Subject
if any of the following circumstances occurs, the investigator must discontinue the study drug administration to the subject immediately, provide him with appropriate treatments and perform the examinations/observations scheduled at the time of discharge as much as possible. Subjects prematurely withdrawn from the study may be replaced. The investigator will document the date of and reason for discontinuation, treatments given after discontinuation and clinical course thereafter in the CRF, which must be submitted to the sponsor without delay. If the scheduled examinations/observations were not performed at discontinuation, the reason must be documented in the CRF.
| Number | Date | Country | |
|---|---|---|---|
| 62431014 | Dec 2016 | US |
| Number | Date | Country | |
|---|---|---|---|
| Parent | 16467410 | Jun 2019 | US |
| Child | 17506012 | US |
