HUT74275A - Process for treatment of rheumatoid arthritis - Google Patents

Description

The present invention relates to a method of treating patients with some type of chronic inflammatory arthritis, and more particularly, to the use of a corticotropin-releasing factor or analogues to reduce the periodic acute exacerbation of a chronic inflammatory condition.

Chronic inflammatory arthritis is one of the groups of diseases that affect people; these include rheumatoid arthritis (chronic arthritis), Reiters syndrome, ankylosing spondylitis (spondylitis) and inflammation of the bowels. Of these, rheumatoid arthritis causes approximately 33,000 deaths each year in the United States. The disease is currently not curable and the cause is unknown. Patients with rheumatoid arthritis experience intermittent flare-ups, that is, occasionally acute reactions lasting from one week to one month.

Steinman explores some possible new approaches to autoimmune diseases, including rheumatoid arthritis, in Autoimmune Disease (Scientific American, 107-114, September 1993). One of these is the design of trap insertion into the HLA (human lymphocyte antigen) cleavage to turn off T cells. In another approach, a monoclonal antibody that binds to tumor necrosis factor appears to increase joint motility and reduce rigidity due to rheumatoid arthritis. However, these new approaches are not yet well established to be therapeutically useful.

Treatment models using rat strains prone to arthritis are facilitated by treatment research. Lewis rats susceptible to arthritis, among other possible abnormalities, respond to various stimuli with reduced hypothalamic CRF (corticotropin-releasing factor), so researchers can experimentally induce arthritis and some other autoimmune diseases. In the Piebald-Viral-Glaxo (PVG) rat model, animals do not show defects in the hypothalamus-pituitary-adrenal axis other than the Lewis rat strain, but are prone to arthritis.

U.S. Patent No. 5,006,330 to Sternberg et al. Discloses the use of a Lewis rat model in which experimental animals develop arthritis that mimics human rheumatoid arthritis in response to group A of streptococcal cell wall fragments. The teaching teaches the use of the model to investigate the predisposition of mammals to inflammatory diseases. Sternberg et al. Describe the measurement of CRH (corticotropin-releasing hormone) after injection of various reagents. CRH is another name for CRF, a 41-amino acid neuropeptide in the brain and peripheral nerve endings that stimulates the secretion of ACTH from cells in the brain, pituitary glands, and distant peripheral sites such as nerve endings and white blood cells. . Sternberg et al. Also report that CRH injections have been administered after measurement of ACTH and corticosterone.

It has been reported in the literature that antibodies to CRF partially abolish the inflammatory response, and on this basis, various researchers, such as Karalis et al., Have reported that autocrine or paracrine inflammatory actions of Corticotropin-Releasing.

4 Hormones in Vivő (Science, 254: 421-423 (1991)) suggest that CRF is an inflammatory agent.

U.S. Patent No. 4,489,163 to Rivier et al. Discloses rat CRF and its analogs. Human CRF has the same sequence as rat CRF. U.S. Patent No. 4,415,558 to Vale, Jr. et al. Describes the synthesis of sheep CFR (oCRF) and its analogs and the isolation of oCRF from sheep hypothalamic extracts. OCRF has been found to lower blood pressure.

As a generally similar peptide, sauvagine [Regulatory Peptides, 2 <1 ^_ 13 (1981)]. This peptide consists of 40 amino acids and has been reported to have the biological effect of lowering blood pressure in mammals and stimulating the secretion of ACTH and β-endorphin.

In U.S. Patent Nos. 4,528,189 and 4,553,654, Lederis et al. Describe peptides and analogues similar to rat and sheep CRF and found that this analogue, urotensin of rabbit fish and carp, stimulates ACTH. and lowers blood pressure. The CRF-related peptide, the urotensin of rabbit fish, has the same sequence as the carp urotensin, except that it contains isoleucine at position 24 and glutamic acid at position 27.

Ling et al. (BBRC, 122: 1218-1224 (1984)) describe a goat CRF identical to sheep CRF. ESR et al. (BBRC, 122, 899-905 (1984)) disclose the structure of calf CRF that differs from sheep and goat CRF in only one amino acid

···· ·· · ·· ·· (in position 33 here is aspartic acid, while in sheep and goat CRF is serine). Patthy et al., Proc. Natl. Acad. Sci. 82: 8762-8766 (1985)] isolated and characterized porcine CRF. The amino acid sequence of porcine CRF is shown in Figures 1-39. sections are identical to rat and human CRF and differ only in positions 40 and 41. At position 40 it contains aspartic acid or isoleucine and at position 41 it contains phenylalanine amide.

Wei, U.S. Patent No. 4,801,612, discloses that an (acute) inflammatory reaction to the skin or mucosa is inhibited by administering to the patient a corticotropin-releasing factor or an analogue thereof, and in U.S. Patent No. 5,137,871 or U.S. Pat. salt or analogue) for use in treating an injury or disease of the brain, nervous system, or muscles where edema may occur.

Adding a corticotropin-releasing factor to a patient with a type of chronic inflammatory arthritis, such as rheumatoid arthritis, reduces the risk of intermittent acute inflammatory reactions. CRF, when used in the dosages of the present invention, exhibits anti-inflammatory activity in the advanced phase of the disease in the form of a dose response. Thus, a further object of the present invention is to provide a therapeutic method for the treatment of these persistent diseases which cannot be controlled by current therapies.

Treatment according to the invention must be performed under the supervision of a health care professional, usually an internist, and coordinated with other medications or treatments. Since the treatment of acute inflammatory reactions ···· ··, which usually takes from one week to several months, results in anti-inflammatory effects and other improvement results within a few days, patients may have an unpleasant or they experience a significant reduction in the symptoms of incapacitation, although the use of the invention is not a cure.

BRIEF DESCRIPTION OF THE DRAWINGS Fig. 1 is a perspective view of two Lewis LEW / N rat paws susceptible to myobacterial-induced arthritis in which 10 paws are a control sample and 12 paws are injected with Myobacterium to induce arthritis;

Figure 2 shows the swelling of the sole (unit = 0.025 mm) versus time in an animal model of arthritis-prone rats in control animals and animals treated with the method of the invention (50 and 100 pg / kg subcutaneous CRF injection) where the swelling of the sole was measured in the (left posterior) paw injected with myobacteria;

Fig. 3 is similar to Fig. 2, except that measurements are taken on paws (opposite posterior) opposite to the injected paws;

Figure 4 is a graph showing the estimated severity of arthritis over time in the right hind paw treated as shown in Figure 3;

Figure 5 is similar to Figure 4 except for the other two paws (left and right first);

Fig. 6, which includes sub-drawings A-E, is similar to a

2, only here with the method according to the invention a higher dose

7 (100 µg subcutaneously injected CRF in the left hind paw injected with myobacterium) with a delay of up to 40 days after disease induction;

Figure 7, which includes A-E sub-drawings, is similar to Figure 6 except for measurements on transverse (right posterior) paws; and Figure 8 is a graphical representation of the occurrence of arthritis, similar to Figure 4, but using the treatment regimen described in Figure 6.

Preferred embodiments of the invention will now be described in detail.

Earlier research reports that CRF is present in the joints of mammals with inflammatory arthritis, and consequently that CRF is an inflammatory agent, may need to be corrected because the peptide may also be an inflammatory agent. how much it is produced endogenously, how the inflammatory cell reacts to CRF, and when CRF appears during the inflammatory reaction. The following research description demonstrates that the administration of CRF may exert a dose-response therapeutic effect in the inhibition of intermittent acute and chronic inflammatory reactions, so that the present invention is a therapeutic method for treating patients with some type of chronic inflammatory arthritis such as rheumatoid arthritis.

CRF is used in the process of the invention. CRF also includes known analogs of CRF and related peptides. THE

You can make CRF-ready pharmaceuticals for storage • · · · * · · · · · ·

8) in the form of lyophilized masses or aqueous solutions, optionally mixed with CRF of desired purity with physiologically acceptable carriers, excipients or stabilizers. Acceptable carriers, excipients, and stabilizers at the dosages and concentrations employed are non-toxic to the patient and include buffers such as phosphate, citrate or other organic acids; antioxidants such as ascorbic acid; low molecular weight (less than about 10 amino acids) polypeptides; proteins such as serum albumin, gelatin or immunoglobulins. Additional components may include glycine, glutamine, aspartic acid, arginine or lysine; monosaccharides, disaccharides and other carbohydrates such as glucose, mannose, 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 of the known methods, for example, intra-articular (articular), intravenous, intrathecal (intravaginal), subcutaneous injection, intranasal (nasal), orally or via an implant. Gel foam, wax or microparticulate formulations may be used as implants. Topical formulations preferably contain transdermal delivery aids, such as solubilizing agents or guided delivery systems, which are known or are in the process of being developed in the context of rapidly developing skin patches.

When using the method of the invention, CRF or • ·

The therapeutically effective dose of its 9 analogs to humans is preferably at least 0.01 µg / kg. For systemic (e.g., subcutaneous or intravenous) administration, doses are more preferably from about 0.1 to about 50 µg / kg, most preferably from about 1 to about 30 µg / kg. For topical (e.g., intra-articular) administration, a dosage of about 1 to about 100 mg / kg is preferred. Doses of CRF may be administered by slow infusion, for example subcutaneously or intradermally, or injected directly into the affected body.

The treated patient should be supervised by a health care professional, usually a physician, or the physician may self-administer or prescribe the CRF-containing medicinal product (eg, with a pre-filled syringe). Systemic (general-purpose) modes of administration are particularly preferred, as intermittent acute situations or flares often occur at several sites simultaneously. The treatments of the invention, as illustrated in more detail below, are effective, but unfortunately, the effect is reduced when treatment is complete. As with many other treatments, the method of treatment of the invention may be combined with other drugs and other therapeutic approaches to the disease being treated.

CRF can be administered according to the invention for about three weeks, but side effects (such as Cushing's syndrome) may occur with prolonged use. Thus, the present invention is primarily directed to the treatment of acute intermittent flares, and CRF is administered for up to 40 days from the onset of such flares.

Although typically soluble in water, typically, the peptides may be administered in the form of their pharmaceutically acceptable non-toxic salts, such as acid addition salts such as hydrochloride, hydrobromide, sulfate, acetate, citrate, benzoate, succinate, maleate, ascorbate, tartrate. In addition to being administered with any pharmaceutically acceptable carrier, CRF may be processed into a liquid carrier such as liposomes, microcapsules, polymer or wax-based and controlled release formulations, or made into tablets, pills, or capsules.

The method of treatment of the present invention is applicable to patients with some type of chronic inflammatory arthritis. This group of diseases includes rheumatoid arthritis, Reiters syndrome, ankylosing spondylitis, psoriatic arthropatica (psoriatic arthritis) and inflammation of the gut. The treatments are particularly useful for intermittent acute flares and provide an anti-inflammatory effect in the form of a dose response. Experiments show that the method of the present invention has anti-inflammatory activity, even during the advanced phase of the disease, regardless of the duration of application (e.g., up to about 40 days from the start of the intermittent acute flare-up), but inflammation symptoms reappear when treatment is discontinued. Therefore, the present invention is not a curative but rather a treatment method.

Example

The therapeutic effect is shown in a rat model of adjuvant arthritis in which Mycobacterium killed by arthritis was injected into the left hind paw of male Lewis LEW / N rats.

In Figure 1, Figure 10 shows the paws of saline-treated animals 16 days after disease induction, while the paws labeled 12 with mycobacterium (and later, as will be apparent from the description, the other paws) swelling of the sole was observed. Swelling of the soles is a sign of inflammation. As we know, inflammation is a reaction of the body to injury or attack and takes the form of redness, swelling, warmth and pain. Five days after disease induction, CRF was administered subcutaneously twice daily at doses of 50 Atg / kg and 100 µg / kg, respectively. Twice daily dosing was continued for 20 days. In CRF-treated animals, swelling and inflammation were relatively low by about 16 days after induction.

Swelling was determined by measuring the thickness of the left hind (injected) paw starting on the first day of CRF administration (i.e., 5 days after injection with Mycobacterium). This measurement was used to determine the degree of primary or acute inflammation. Then (usually for about 10-12 days), the right hind paw was also swollen and inflamed. This is a chronic inflammatory reaction.

Figure 2 shows swelling data for the primary inflamed left paw over a period of 32 days. These were measured on the one hand in control rats and on the other hand in rats receiving CRF 50 and 100 µg / kg twice daily, respectively. One unit on the vertical axis is 0.025 mm.

As shown in Figure 2, the administration of CRF according to the present invention has a dose-response-like anti-inflammatory effect. In the figure, each curve 3 begins 5 days after injection of Mycobacterium into the left hind paw. Swelling of 50 mg / kg / day resulted in less swelling compared to the comparator, and 100 mg / kg of CRF had an even greater effect.

The data in Figure 3 is comparable to that in Figure 2, but the swelling of the sole is measured here on the right hind paw, which is a chronic inflammatory reaction. Dosing of 50 and 100 IU / kg CRF was started here, as in Figure 2, 5 days after disease induction, but the thickness of the right hind paw began to increase later than that of the left hind paw in Figure 2. The swelling of the right hind paw is also smaller than that of the left hind paw. Of course, the right hind paw was not affected as directly as the left hind paw. However, after about 11 days, the right hind paw will begin to swell; however, animals that received CRF (up to 50 or 100 IU / kg) according to the present invention showed very little swelling.

In the animals treated as shown in Fig. 3, the extent of arthritis was determined by a scoring method in which 0 denotes no arthritis and 4 denotes severe arthritis; the results are shown in Figure 4. Arthritis scores were calculated by taking into account the percentage of sole involvement with the general symptoms of the disease (swelling, redness and distortion). The results shown in Figure 4 show that both doses of CRF (50 and 100 mg / kg) reduce arthritis scores, but the 100 µg / kg dose is more effective. The scoring was used to assess the right hind paw of the animals.

In Figure 5, similar to Figure 4 above, the two forelegs were examined. Thus, Figures 4 and 5 show a chronic rather than an acute situation. For the first paws, the results of which are shown in Figure 5, the increase in iron throat and other symptoms were delayed even more, and were milder than the symptoms in the right hind paws shown in Figure 4.

2-5. In the experiments summarized in Figures 1A to 5C, treatment started 5 days after disease induction. In another series of experiments, the disease was induced as before and the animals were treated with 100 µg / kg subcutaneously twice daily in the manner of the invention, except that the treatment after 5, 10 , 15, 20, 25 and 40 days, respectively. The results are summarized in Figures 6-8. is shown.

Figure A-E in Figure 6 shows measurements on the left hind foot. Figure A shows the thickness of the sole in control animals and those treated from day 5 (indented with arrow) to day 20 (indented) with disease induction. Immediately after commencement of treatment according to the invention on day 5 following the induction of disease, the swelling of the sole was significantly reduced compared to the control, and this reduction continued until treatment was discontinued; then the swelling started and it lasted for about 60 days by which time it was • ·

There was practically no difference between the previously treated and the control (i.e., untreated) soles. Similarly, in Figure B) the data of control and treated rats from day 10 to day 30, in figure C) from day 15 to day 35, in D) from day 25 to day 45 and Figure E shows data for rats treated from day 40 to day 60. These results indicate that CRF treatment greatly reduced the swelling of the soles, even in the advanced phase of the disease, as shown in Figure E). Figure 6 (E) shows that although treatment began only on day 40 after disease induction, swelling on the treated sole immediately decreased upon initiation of treatment.

Figures A-E of Figure 7 are similar to those of Figure 6 except that the swelling was measured on the right hind foot. That is, as in Figure 3, despite the fact that the measured data refer to the right hind foot, while injecting mycobacterium in the left hind foot (thus representing an acute flare), there may be chronic inflammatory reactions in the right hind foot. . Again, the results show that when CRF is administered according to the invention, it significantly reduces swelling in chronic inflammation and even in the advanced phase of the disease.

In experiments similar to those of Figures 6 and 7, in which the degree of arthritis was determined as described in Figure 4, the method of the present invention produced a significant effect on the general symptoms of arthritis, albeit somewhat less than that observed in reducing foot swelling. In the drawings A-E in Fig. 8, treated animals exhibit

- The beginning of 15 treatments was marked with an empty arrow and the end of treatment with a filled arrow.

In the experiment of Figure 8A, five days after the injection of mycobacterium into the left hind paw of an animal, CRF was administered to the right hind paw according to the invention for twenty days. During treatment, arthritis scores decreased relative to the control sample, but when treatment was discontinued, scores increased. Thus, the method of the invention is not prophylactic but therapeutic. Importantly, however, as shown in Figure 8D, the treatment of the present invention is effective when applied only on the 25th day of the disease.

The foregoing has been described in connection with the preferred embodiments of the invention, but the description and the examples are provided by way of illustration only and are not intended to limit the scope thereof.

Claims (11)

PATENT CLAIMS CLAIMS 1. A method of treating patients with some type of chronic inflammatory arthritis, comprising administering to the patient CRF at therapeutically effective doses for at least a few days. 2. The method of claim 1, wherein the method is for treating patients with rheumatoid arthritis. The method of claim 1, wherein each dose administered is at least 0.01 µg / kg body weight. The method of claim 1, wherein the administration is by infusion or absorption. The method of claim 1, wherein the administration is by topical injection, each at a dose of about 1 to about 100 µg / kg body weight. 6. A method of treating patients with rheumatoid arthritis, comprising administering to the patient a therapeutically effective dose of CRF over a period of at least a few days in an amount effective to reduce inflammation of a affected body. 7. The method of claim 6, wherein the administration reduces the patient's intermittent acute inflammatory reactions. 8. The method of claim 6, wherein the administration is by topical, intra-articular, intravenous, intrathecal, subcutaneous injection, orally, intranasally or by means of an implanted device. 9. The method of claim 7, wherein administration is initiated within about forty days after intermittent acute inflammation. 10. A 9th According to claim 1 live flood, with well analyzed, that treatment at most about three each other the following week through to be continued. 11. A 8th According to claim 1 live flood, with well analyzed, that each dose ranges from about 0.1 to about 50 µg / kg for systemic administration and from about 1 to about 100 µg / kg for topical administration. Process according to claim 1 or 6, characterized in that CRF is used in the form of its salt. 13. The method of claim 1 or 6, wherein the CRF is used in an analog form.