CZ122696A3 - The use of crf corticotropin releasing factor or analogs thereof for preparing a therapeutical preparation - Google Patents

Abstract

Administration of corticotropin-releasing factor to patients suffering from one of the chronic inflammatory arthritides, such as rheumatoid arthritis, reduces episodic acute inflammatory responses to the chronic disease. Doses of administered CRF in accordance with the invention give anti-inflammatory results in a dose responsive manner even in well-established disease.

Description

Technical field o

The invention relates generally to a method of treating a patient suffering from chronic inflammatory arthritis, and more particularly to the use of a corticotropin release factor or analogues thereof to reduce swelling during episodic acute reactions in chronic inflammatory conditions.

BACKGROUND OF THE INVENTION

Diseases affecting mankind include chronic inflammatory arthritis, including rheumatoid arthritis, Reiters syndrome, ankylosing spondylitis, psoriatic arthritis, and inflammatory bowel disease. It is estimated that rheumatoid arthritis alone causes about 33,000 deaths in the United States each year. There is no cure for this disease yet and its cause is unclear. Patients with rheumatoid arthritis periodically experience flares or episodic acute reactions lasting about a week to about a month.

Steinman, in "Autoimmune Disease, Scientific American, pp. 107-114 (September 1993), discusses some possible new approaches to the treatment of autoimmune disease, such as rheumatoid arthritis. In one of these, it is proposed to knock out T cells using dummies that fit into the HLA (human lymphocyte antigen) cleft. Furthermore, a monoclonal antibody that binds to tumor necrosis factor appears to increase joint mobility and reduce stiffness associated with rheumatoid arthritis. However, these possible novel approaches are not yet therapeutically stabilized in terms of efficacy.

The search for a possible treatment is aided by the fact that animal models have been developed using rat susceptible to arthritis. Among other possible abnormalities, Lewis-susceptible arthritis rats have a deficient hypothalamic corticoliberin response to various stimuli, so researchers can experimentally induce inflammatory arthritis and some other autoimmune diseases. In the rat model of Piebald-ViralGlaxo ("PVG"), unlike Lewis rats, animals do not appear to have a defect in the hypothalamic-pituitary-adrenal ("HPA") axis, but are susceptible to arthritis.

U.S. Patent No. 5,006,330 to Sternberg et al., Issued April 9, 1991, describes the use of an animal model of Lewis rats where experimental animals developed arthritis that simulates human rheumatoid arthritis in response to exposure to cell wall fragments of group A. Sternberg et al. use of this model to test the susceptibility of mammals to inflammatory diseases. Sternberg et al. Describe the measurement of CRH (corticotropin releasing hormone) after injections of various formulations. CRH is an alternative name to corticoliberin (corticotropin discharging factor, CRF), a 41 amino acid neuropeptide that is present in the brain and peripheral nerve endings and stimulates the release of ACTH from pituitary cells in the brain and from various peripheral sites such as are nerve endings and leukocytes. Sternberg et al. Also describe CRH injections and subsequent ACTH and corticosterone measurements.

It has been reported that the inflammatory response is partially abolished by antibodies directed against CRF, leading various researchers to believe that CRF is a pro-inflammatory agent. Karalis et al., "Autocrine or Paracrine Inflammatory Actions of Corticotropin-Releasing Hormone In Vivo, Science 254, 421423 (1991).

U.S. Patent 4,489,163, Rivier et al., Issued December 18, 1984, describes rat CRF and analogs thereof. Human CRF has the same sequence as rat CRF. Several CRF analogs are known in the art. U.S. Patent No. 4,415,558 to Vale Jr. and d., issued November 15, 1983, describe the synthesis of sheep CRF, analogs thereof, and isolation of oCRF from sheep hypothalamic extracts. Synthetic oCRF has been found to lower blood pressure.

A generally similar peptide of sauvagine has been described in Regulators of Peptides 2, pp. 1-13 (1981). Sauvagine is a 40 amino acid peptide and has been reported to have a biological effect in lowering blood pressure in mammals and stimulating secretion of ACTH and β-endorphin.

U.S. Patent 4,528,185 issued to Lederis et al

And U.S. Patent 4,533,654 issued to Lederis et al

On Aug. 6, 1985, disclose rat and sheep CRF-like peptides and their shieldon analogs, respectively.

disclose that this urotensin carp stimulates ACTH and lowers blood pressure. The CRF-related peptide, i.e. urotensin shield bearer, has the same amino acid sequence as carp urotensin except that at position 24 the amino acid isoleucine and at position 27 the glutamic acid.

Ling et al., BBRC 122, pp. 1218-1224 (1984), disclose a structure of goat CRF that is the same as sheep CRF. Esch et al., BBRC 122, pp. 899-905 (1984) disclose a bovine CRF structure that differs from sheep and goat CRFs in only one amino acid residue (at position 33 there is asparagine instead of lilac, which is at position 33 u goat and sheep CRF). Pig CRF was isolated and characterized by Patthy et al., Proc. Nati. Acad. Sel 82: 8762-8766 (1985). Porcine CRF shares a common amino acid sequence (residues 1-39) with rat / human CRF and differs only at positions 40 and 41.

The residue 40 can be either asparagine or isoleucine and the residue is phenylalaninamide.

U.S. Patent No. 4,801,612 to Wei, issued January 31, 1989, discloses inhibiting an inflammatory response (acute) to the skin or mucous membranes of a patient by administering CRF or analogs thereof, and U.S. Patent No. 5,137,871 to Wei, issued August 11, 1992, discloses the use of CRF (or salt or analogue) in the treatment of a patient with damage or illness of the brain, nervous system or musculature where swelling is a factor.

SUMMARY OF THE INVENTION

Administration of a corticotropin discharge factor to patients suffering from one of the chronic inflammatory arthritis, such as rheumatoid arthritis, reduces the episodic acute inflammatory response to the disease. The doses of CRF of the invention provide anti-inflammatory results in the form of a dose response even in a stabilized disease. Thus, the invention provides therapeutic treatment for these non-remitting diseases that do not address conventional therapies.

The therapeutic treatment of the invention should be performed under the supervision of a health care professional, usually a physician, and may be conducted in conjunction with other drugs and therapies. Since the treatment of episodic acute inflammatory reactions, which would typically last for weeks to months, has been reported to have an anti-inflammatory effect and other favorable results after only a few days, the patient is effectively relieved of unpleasant or even incapacitating symptoms during treatment. although the practice of the invention is not a treatment.

It is possible that earlier reports that detected the presence of CRF in the joints of mammals with inflammatory arthritis and concluded that CRF is a pro-inflammatory agent might be right because it is possible that this peptide may be either pro-inflammatory or anti-inflammatory depending how much endogenous peptide is produced, how “an inflammatory cell responds to CRF, or when CRF occurs during an inflammatory response. However, the following description demonstrates that administration of CRF may be therapeutically effective in inhibiting episodic acute and chronic inflammatory responses in the form of a dose response, and the invention therefore provides a therapeutic method of treating patients with chronic inflammatory arthritis such as rheumatoid arthritis.

CRF is used in the practice of the invention. CRF includes CRF-related analogs and peptides known in the art. Therapeutic CRF formulations may be formulated for storage by mixing CRF of the desired degree of purity with optional physiologically acceptable carriers, vehicles or stabilizers, in the form of a lyophilized cake or aqueous solutions. Acceptable carriers, vehicles, or stabilizers are nontoxic to the recipient at dosages and concentrations used in the administration and include buffers such as phosphate, citrate and other organic acids, antioxidants including ascorbic acid, low molecular weight (less than about 10 residues) polypeptides, proteins such as serum albumin, gelatin or immunoglobulins. Other ingredients may include glycine, glutamine, asparagine, arginine or lysine, monosaccharides, disaccharides and other carbohydrates including glucose, mannose or dextrins, chelating agents such as EDTA, sugar alcohols such as mannitol or sorbitol, salt-forming counterions such as sodium , and / or nonionic surfactants such as TWEEN, PLURONICS or PEG.

Administration can be by any known means, for intraarticular, subcutaneous, injectable, including, intravenous, intranasal, but not intrathecal, oral, or implantable device administration. Suitable implants include, for example, gel foam, wax, or microparticle-based implants. The topical application preferably comprises means or devices for facilitating the delivery to the skin, such as solubilizing or controlled delivery means, which are known and are being developed in the rapidly emerging technologies of the skin delivery systems.

Therapeutically effective doses of CRF or analogs thereof for practicing the invention are preferably at least about 0.01 pg / kg in humans. For systemic administration (e.g., subcutaneous and intravenous), dosages in the range of about 0.1 to about 50 pg / kg, and more preferably about 1 to about 30 pg / kg, are more preferred. For topical administration (e.g., intra-articular), a dose in the range of about 1 to about 100 μς / kg is preferred. CRF doses may be administered slowly, for example, subcutaneously or intradermally, or may be injected directly into the affected part of the body.

The patients to be treated will be in the care of a healthcare professional, such as a physician, who can self-administer a therapeutic product comprising CRF or prescribe it to a patient for self-administration (such as pre-filled syringes). Systemic dosage forms are particularly advantageous in view of the frequent multiple occurrence of episodic situations or disease outbreaks. Although the treatment according to the invention has been shown to be effective, as explained below, it is unfortunately also discontinued when treatment is interrupted. As with many other types of therapies, the therapeutic method of the invention may be combined with other pharmaceuticals and other therapeutic approaches to the disease being treated.

The CRF of the invention may potentially be continued for up to about three weeks, but side effects (e.g., Cushing's syndrome) may develop upon prolonged intake. Thus, application of the invention is primarily contemplated for the treatment of acute, episodic outbreaks of the disease and administration of CRF may be initiated up to forty days after the appearance of episodic acute inflammation.

Although peptides as typically synthesized,. are generally water-soluble, and can be administered in the form of pharmaceutically acceptable non-toxic salts, such as acid addition salts. Examples of acid addition salts include hydrochloride, hydrobromide, sulfate, acetate, citrate, benzoate, succinate, malate, ascorbate, tartrate and the like. In addition, CRF may be administered in any pharmaceutically acceptable carrier, depending on the desired The method of administration may be formulated together with the liquid carrier in controlled release liposomes, microcapsules, polymers or wax-based formulations, or may be formulated in the form of tablets, pills or capsules.

The method of treating a patient of the invention relates to patients suffering from one of the chronic inflammatory arthritis. Chronic inflammatory arthritis herein includes rheumatoid arthritis, Reiters syndrome, ankylosing spondylitis, psoriatic arthritis, and inflammatory bowel disease. The treatment is particularly suitable for episodic acute inflammation and provides anti-inflammatory results in the form of a dose response. Described experiments show that irrespective of the time of administration (for example, up to about forty days after episodic acute inflammation) in the practice of the invention, inflammation is also reduced in a stabilized disease; however, if treatment is discontinued, symptoms have been found to recur. Thus, the invention is a treatment rather than a treatment.

BRIEF DESCRIPTION OF THE DRAWINGS

Giant. 1 is a perspective view of two legs of male Lewis " LEW / N " rats in which mycobacterial susceptibility to arthritis has been established, with leg 10 being control and leg 12 being injected with mycobacteria to induce arthritis.

Giant. 2 graphically depicts paw swelling (one unit is equivalent to 0.001 inch) in an arthritis-prone rat animal model versus time for control animals and animals treated according to the invention with two different doses (subcutaneous injection of 50 and 100 µg CRF per kg body weight) , wherein the paw swelling is measured on mycobacteria-injected legs (left hind).

Giant. 3 is analogous to FIG. 2, but shows measurements on the legs in a transverse plane opposite the injected legs (right hind).

Giant. 4 graphically depicts the time-dependence of arthritis for the right hind leg of FIG. 3.

Giant. 5 is analogous to FIG. 4, but relates to a second pair of legs (left and right front).

Giant. 6, comprising panels A to E, is analogous to FIG. 2, but shows a larger dose (100 µg / kg body weight) administered subcutaneously to a paw injected with mycobacteria (left hind), according to the invention, from times delayed to 40 days after the onset of the disease.

Giant. 7, comprising panels A to E, is analogous to FIG. 6, but relates to the opposite paw (right rear).

Giant. 8 graphically depicts the dependence of arthritis in a manner similar to FIG. 4 but in a similar format to FIG. 6.

DETAILED DESCRIPTION OF THE INVENTION

Therapeutic efficacy was demonstrated in a rat model of adjuvant arthritis in which male Lewis rats "LEW / N, susceptible to arthritis, were injected into the left hind paw of killed mycobacteria to induce arthritis.

Referring generally to Fig. 1, the animals (i.e., saline-treated animals) had their legs shown as foot 10 approximately sixteen days after the disease was induced; however, shortly after disease induction, mycobacteria-treated legs 12 (and later, as described, other legs) experience paw swelling. Paw swelling is an inflammatory signal. As is known, inflammation is manifested by redness, swelling, heat and pain as a reaction of the body against injury or attack. Five days after disease induction in rats, CRF was administered subcutaneously twice daily at either 50 µg / kg or 100 µg / kg single dose. This twice daily administration was continued for 20 days. At approximately day 16 after disease induction, CRF treated animals had relatively less swelling or inflammation.

Beginning on the first day of administration (i.e., day 5 after injection of killed mycobacteria), the magnitude of the swelling was determined by calipers by measuring the thickness of the left hind (injected) leg. This measurement served to determine the degree of "primary or acute inflammation." Subsequently (usually after about another 10 to days) it also swells and the right hind leg also becomes inflamed. This is a chronic inflammatory reaction.

Figure 2 shows swelling data for the primary inflamed left hind leg (measured until day 32) for control rats, for rats receiving 50 CRF body weight twice daily, and for rats receiving twice daily dosing.100 μρ / kg body weight CRF. The vertical axis is indicated by units, one of which is 0.001 inch. As can be seen from the data in Figure 2, administration of the CRF of the invention gave anti-inflammatory results in the form of a dose response. All three plots in Figure 2 start 5 days after challenge with mycobacteria. At 50 µg / kg CRF injection twice daily, swelling was reduced compared to control. At 100 μρ / kg, the results were even more significant.

Fig. 3 shows analogous data to Fig. 2, but the measured degree of paw swelling relates to the right hind leg and thus represents a chronic inflammatory reaction. CRF dosing at either 50 μς / kg or 100 μς / kg was started 5 days after the challenge as in Fig. 2, but the right hind leg had swelling later than the left, for which data is shown in Fig. 2. The right hind leg also exhibits less swelling than the left hind leg. Of course, the right hind leg is not directly provoked as the left leg. However, starting from day 11, the right hind leg begins to swell; however, in the case of animals receiving the CRFs according to the invention, a very small swelling (at both 50 μg / kg and 100 μg / kg body weight) is reported.

As shown in FIG. 4, the extent of arthritis in the animals shown in FIG. 3 was evaluated by a scoring system in which “0 represents no arthritis and“ 4 represents severe arthritis. Arthritis assessment was performed based on a combination of factors including the percentage of paw edema plus the overall symptoms of the disease (which include swelling, redness, and disfigurement). The data in Fig. 4 show that both doses of CRF (50 μς / kg and 100 were effective in reducing the symptoms of arthritis, but appear to have a greater effect at the dose of 100 μς / kg). animal feet.

In Fig. 5, both front legs were evaluated analogously to Fig. 4. Giant. 4 and 5 therefore represent chronic rather than acute conditions. However, the anterior legs used for the data in Fig. 5 experienced even more delayed and less pronounced onset of swelling and other symptoms than the right hind leg of Fig. 4. As can be seen in particular from Fig. 5, CRF administration according to the invention is very effective. in the treatment of clinical manifestations of chronic inflammatory arthritis.

In the experiments whose data are summarized in Figures 2 to 5, treatment was started 5 days after disease induction. Another set of experiments was performed where the disease was induced as before and the animals were treated according to the invention (with a subcutaneous dose of 100 μρ / kg twice daily) for 20 days except that the course of treatment began 5 days, 10 days, 15 days, 25 days. days or 40 days after the disease is induced. Data from these experiments are summarized in Figures 6 to 8.

In Fig. 6 panels A to E show measurements taken on the left hind paw. In panel A, the paw thickness is plotted for control rats and for rats receiving treatment starting on day 5 after disease induction (indicated by an empty arrow) and ending on day 20 (indicated by a solid arrow). Thus, starting just after the treatment of the invention 5 days after disease induction, the paw thickness was greatly reduced compared to control and the thickness reduction continued until treatment was discontinued when swelling began and continued until about day 60 when there was essentially no difference between pretreated paw and control. (never treated) paw. Similarly, panel B shows data for control and test rats at start of treatment at day 10 and end at day 30, panel C at start of treatment at day 15 and end at day 35, panel D at start of treatment at day 25 and end at day 45 and panel E starting treatment at day 40 and ending at day 60. These data demonstrate that in CRF treatment, paw edema is dramatically reduced even in the stabilized disease shown in panel E. Thus, in panel E in Figure 6, treatment began 40 days after induction. disease, but immediately after the start of treatment, the treated paw decreased in swelling.

In Fig. 7 panels A to E show analogous data to Fig. 6, but the paw swelling was measured on the right hind leg of the animal. That is, as in Fig. 3, since the degree of paw edema was measured on the right hind leg (where the mycobacteria injection site is the left hind leg, which may represent an acute flare), the right leg may represent chronic inflammatory reactions. Once again, these data demonstrate that administration of the CRF of the invention in the case of chronic inflammatory conditions markedly reduces swelling even in stabilized disease.

In experiments similar to Figures 6 and 7, however, where the extent of arthritis in animals was assessed by the system described for Figure 4, the practice of the invention has a significant effect against the overall arthritic condition, albeit less than the apparent decrease in paw edema. In Fig. 8, treated animals are shown in each of panels A to E, wherein the start time of treatment is indicated by empty arrows. The treatment end time is indicated by solid arrows.

Referring to panel A of Figure 8, i.e. 5 days after provoking the left hind paw of the animal by killed mycobacteria, administration of the CRF of the invention to the right hind leg was started and continued for 20 days. During treatment, arthritis scores decreased compared to controls, but increased again after treatment was discontinued. Thus, the practice of the invention is therapeutic, not prophylactic. However, it is important to note that, as shown in panel D of Figure 8, initiation of the treatment of the invention after 25 days of disease is still effective.

The invention, which has been described herein in connection with the preferred specific embodiments, is to be understood that the description and examples are given only for illumination and not for limiting the scope of the invention, which is defined by the appended claims.

Claims (2)

PATENT CLAIMS Use of corticotropin releasing factor CRF or an analogue thereof for the preparation of a therapeutic agent for the treatment of chronic inflammatory arthritis. Use according to claim 1 for the preparation of a therapeutic composition for the treatment of rheumatoid arthritis. Use according to claim 1, wherein the CRF is in the form of a salt. Use according to claim 1, wherein a CRF analog is used.