JPH037237A - Novel use and therpeutic means for administering cyclospoline - Google Patents

Abstract

PURPOSE: To treat intestinal diseases without harmful side-effect by topically dosing an effective amount of cyclosporin into intestines. CONSTITUTION: Cyclosporin is dosed in the intestinal tracts using an arbitrary dosing form topically attainable to the inner face of the tracts to treat intestinal autoimmune diseases such as inflammatory intestinal diseases, ulcerative colitis, Crohn's disease, rectitis, sigmoiditis, complex proctsigmoiditis. The administration form is preferably enema and the dose is 0.5-15.0mg/kg, preferably 2.0-8.0mg/kg, and a therapeutic effect is effectively obtained at a dose level that a high concentration of cyclosporin is not or substantially not observed to infiltrate into tissues.

Description

DETAILED DESCRIPTION OF THE INVENTION [Industrial Application Field] This invention relates to new uses for cyclosporins, particularly new pharmaceutical uses and new methods of pharmaceutical administration of cyclosporins, such as for the treatment of the diseases and conditions described below. .

[Conventional technology]

Cyclosporins are cyclic poly-N-methylated undecapeptides with a unique structure that generally have pharmacological, particularly immunosuppressive, anti-inflammatory and/or anti-parasitic activity, to varying degrees. Includes types of The first cyclosporins isolated were also known as ``cyclosporin A'' and are now under the registered trademark SANDIMUNE (SANDI).
A natural fungal metabolite [cyclosvorine J (Cic
losporin or cyclosporine). "Cyclosporin" is a compound of the formula (A) [wherein, -M
eBmt- has the formula (B) CH(R) CI(.

CHl (wherein -X-Y- is trans-〇H=CH-,
A is -αAbu-) is a cyclosporine represented by N-methyl-(4R)-4-but-2E-en-1-yl-4-methyl-(L)threonyl residue; .

As the "parent" compound of this class, "cyclosporine" has received the most attention to date. The main area of clinical research on Cyclosporin is its application as an immunosuppressant, especially in organ recipients such as heart, lung, combined cardiopulmonary, liver, kidney, spleen, bone marrow, skin and corneal transplants and especially atypical organ transplants. It was there. In this field, ``Cyclosporin'' has achieved remarkable results and has been well received, and is currently commercially available under the registered trademark SAJJDIMMUN.
It is available as a drug and is widely used clinically.

At the same time, "cyclosporine" is used for various autoimmune diseases and inflammatory conditions, especially those with an etiology that includes an autoimmune component, such as arthritis (e.g. rheumatoid arthritis, chronic progressive arthritis and degenerative arthritis) and rheumatoid arthritis. Because of its strong applicability, reports and achievements in vitro, in animal models, and in clinical trials have been extensively described in the literature. Specific autoimmune diseases for which cyclosporine therapy has been proposed or applied include autoimmune hematological diseases (including, for example, hemolytic anemia, aplastic anemia, erythroblastic fistula and idiopathic thrombocytopenia);
Systemic erythematous lupus, multiple sclerosis, scleroderma
odoma), Begener granulocytosis, dermatomyositis, chronic active hepatitis, myasthenia gravis, axial, Steven-Johnson syndrome, idiopathic sprue, autoimmune inflammatory bowel disease (including ulcerative colitis and Crohn's disease) , endocrine opthalmopathy, severe thyroid dysfunction, psychoidosis, multiple sclerosis, primary biliary cirrhosis, juvenile diabetes (diabetes type I), uvea (anterior and posterior), sicca conjunctivitis and spring sicca Conjunctivitis, interstitial pulmonary fibrosis, axial arthritis, and glomerulonephritis (with or without nephrotic syndrome,
For example, idiopathic nephrotic syndrome or lymphoid nephrosis).

Another area of research could be used to treat malaria, schistosomiasis and coccidiomycosis, and to reduce the resistance of other anti-adenomatous or other cytostatic therapies, such as cancer treatments. It has potential applications as an anti-parasitic agent, especially as an antiprotozoal agent.

Since the first discovery of "cyclosporin," a wide variety of natural cyclosporins have been isolated and identified, and many more non-natural cyclosporins have been produced by entirely or semi-synthetic means or by the application of modified culture techniques. That is, there are currently many types of cyclosporins, such as natural cyclosporins A-Z [Reference, Traver et al.
, Acta), Vol. 60, pp. 1247-1.255 (19
1977), Traver et al., 2, “Helvetica stain force.
Actor J(I(elv, chim.

Acta), Volume 65, No. 162, 1655-1667
(1982), Kobel et al., European Journal of Applied Microbiology and Biotechnology J (Europ.

J. Applied Microbiology a.
nd Biotechnology), Volume 14, 273
-240 pages (1982) and Fono-Wartburg et al.
ss in Allergy), Vol. 38, pp. 28-45 (1986) and various non-natural cyclosporin derivatives and dihydro-cyclosporins [formula (B)
By saturating the -xy portion of the -MeBmt- residue in -X y -= -CH! CH! -, artificial or synthetic cyclosporins, derivatized cyclosporins (e.g., cyclosporin molecule with 3 substituents)
), -M
Cyclosporins in which the eBmt-residue is present in an isomeric configuration (e.g., the configuration across the 6° and 7° positions of the -MeB@j- residue is cis rather than trans) and those developed by e.g. Wenger et al. These include cyclosporins in which a mutant amino acid is introduced at a specific position within the peptide sequence using a fully synthetic manufacturing method for cyclosporins [for example, Traper et al. 1, Traver et al. 2 and Korbel et al., supra, U.S. Pat. No. 4108985 and No. 4
No. 210581, European Patent Publication No. 0034567
No. and No. 0056782, International Patent Publication No. WO3
No. 6102080, Wenger 11 “Trans+pl Block J (Trans+pl.

Proc, ), Volume 15, Supplement, 1:2230 (1983
), Wenger 2, ``Unguevante Hemi, International Edition in English J (
Angew, Chew, Int, Ed,), 24
Vol. 77 (1985) and Wenger 3, [Progress in the Chemistry of Organic
Natural Products J (Progress in
the chemistry of organic
Natural Products), vol. 50, 123
(1986)].

In other words, there are currently many types of cyclosporine, such as [Thr]'-1[Val]'-1(:Nv
a] ' and [N val '' - [N val '
- cyclosporin (also known as cyclosporin C5DSG and M, respectively), [3-0-acetyl-M
eBmt]'-cyclosporin (also known as cyclosporin A acetate), [dihydro-Me
B mt] '-[Vall''-cyclosporine (
(also known as dihydro-cyclosporine D),
[iso-MeBmt]'-[Nva]''-cyclosporin (also known as isocyclosporin G), [
(D)Ser]”-cyclosporine, [Me I le
”-Cyclosporin, [(D)MeVal] - Cyclosporin (also known as Cyclosporin H), [MeA1a]@-Cyclosporin, [(D)Pr
o]”-cyclosporin and the like.

[Following conventional nomenclature for cyclosporins, these definitions are provided throughout this specification (including the claims) by presenting the structure of cyclosporin J (i.e., cyclosporin A). An example showing an amino acid residue present that is different from that present in [(D)PrO]3 indicates that the cyclosporin in question has a -(D)PrO-residue in position 3 rather than an 8ar-residue.
ro-), and then by applying the term "cyclosporin" to denote the characteristic that the remaining residues are "the same as those present in cyclosporingo." Each residue is at position 1 -MeBmt
- or dihydro-MeBmt- residues.

The majority of these new cyclosporins have similar medicinal properties to "cyclosporine" or other useful medicinal properties - e.g. activity to eliminate tumor resistance to cytostatic therapy, and their use as therapeutic agents. Numerous proposals regarding this can be found in the literature.

Despite the tremendous contribution made by cyclosporine, especially in the area of organ transplantation and the treatment of autoimmune and inflammatory diseases, there are difficulties in providing effective and convenient means of administration, and there are no reports. Undesirable side effects, especially nephrotoxic reactions, are a major obstacle to its widespread use or adaptation. That is,
"Cyclosporin" is particularly effective for the treatment of psoriasis, autoimmune or inflammatory diseases of the intestine, such as Crohn's disease, and autoimmune or inflammatory diseases of the eye, such as uveitis or vernal keratoconjunctivitis, for example when administered orally. It has been found that. Oral cyclosporine therapy for the treatment of these diseases is warranted, e.g. in severe and chronic cases, or in cases refractory to other therapies, but in less severe cases or at relatively high doses, e.g. The appropriateness of relying on such therapy is questionable when such therapy is indicated. It is easily determined that the risk of developing side effects outweighs the expected therapeutic effect.

In order to overcome these drawbacks, various proposals have been made for topical application of "cyclosporin".

Application in cream or gel form, for example to the lesion in the treatment of psoriasis, or to the surface of the eye, for example in the treatment of uveitis. However, until now, such methods have had little success. Cyclosporin-containing preparations that are applied topically to the skin surface have achieved some efficacy in the treatment of atopic dermatitis, for example, but no clear effect has been shown so far in psoriasis patients. Apart from oral therapy, injections into the affected area are the only remaining effective means of treating psoriasis. Similarly, formulations applied topically to the eye have been found to be effective, for example in preventing rejection of corneal transplants and in treating diseases to the surface of the eye, but in treating deeper diseases, such as posterior uveitis. No clear effectiveness was shown for this.

[Structure of the Invention] Quite contrary to the negative experience with topical application of cyclosporins, according to the present invention, topical application of cyclosporins, e.g. It was found to be extremely effective.

More surprisingly, although such diseases can be treated with effective therapeutic effects, systemic drug penetration or absorption, i.e., high concentrations of cyclosporine entering body tissues as indicated by e.g. serum or whole blood concentration measurements of cyclosporine, is difficult. ,
It has been found that treatment is possible with no or substantially no dosage levels.

The findings of this invention have great therapeutic significance and advantages.

Traditionally, the mainstay of treatment for intestinal diseases such as lower A cancerous colitis has been the use of steroid enemas, and more recently the use of 5-aminosalicylic acid (5-ASA) enemas. but,
Topical steroid and 5-ASA therapy was rarely tolerated. Furthermore, continued steroid therapy often results in mucosal atrophy. The only alternatives for patients for whom such therapy is ineffective or for some reason impossible are usually prophylaxis and colostomy. However, protectectomies and colostomies are not easily tolerated under all circumstances.

The findings of this invention, particularly the knowledge that effective local therapy is possible while reducing the risk of cyclosporine systemic toxicity, represent a significant alternative to conventional therapy and a significant contribution to the well-being of key patient populations currently in need. Promising therapy can be obtained.

Therefore, as a first aspect, the present invention provides the following.

(A) A method of treating an intestinal disease in a subject in need thereof, comprising locally administering a therapeutically effective amount of cyclosporine into the intestine of the subject.

Diseases to which the method of the invention may be applied include those for which cyclosporine therapy is appropriate, such as those of the types mentioned above and other pathological conditions, particularly those attributable to an etiology involving an adverse immune response, an inflammatory response or a parasitic infestation. or a disease or condition that has it. The methods of this invention can also be applied as a means of reducing tumor resistance to other chemotherapy, such as oral anti-neoplastic or cytostatic therapy.

The methods of the invention are particularly applicable to the treatment of autoimmune diseases and conditions and inflammatory diseases and conditions, especially those having an etiology that includes an autoimmune component.

The methods of the invention are particularly useful for treating intestinal diseases, such as those mentioned above. Bowel diseases of particular interest in the context of this invention include inflammatory bowel disease, Crohn's disease, ulcerative colitis, proctitis, sigmoiditis, and proctococcalitis. Intestinal sites where the teachings of this invention can be applied to treat the aforementioned diseases include, among others, the ileum, especially the terminal ileum, the cecum and colon (including the figure 8 colon), and the rectum.

The method of the invention is particularly applicable to diseases that damage the intestinal lining, such as the intestinal mucosa, such as the ileal or colonic mucosa, such as the autoimmune diseases mentioned above.

In the practice of this invention, cyclosporins can be administered intraintestinal or ureterally using any dosage form that can be delivered locally, such as subdivided or similar dosage forms for enteral administration.

However, according to this invention, cyclosporins can be used in liquid form,
Semi-liquid or other fluid or flowable dosage forms, especially aqueous or other hydrophilic liquid dosage forms or foam IJ
most preferably in the form of a flowable gel or the like. For the treatment of intestinal diseases or conditions, cyclosporine is preferably administered in the form of an enema. I wanted to,
This invention provides the following as another series of aspects.

(B) a method of administering cyclosporine therapy for the treatment of intestinal disease in a subject in need of treatment, comprising administering a therapeutically effective amount of cyclosporine by enema; and (C) administering cyclosporine as an active ingredient in the form of an enema. and (D) an enema means for containing or delivering a pharmaceutical composition, the pharmaceutical composition comprising cyclosporine as an active ingredient.

Diseases that can be treated according to (B) above are, for example, those mentioned above. defined as (C) above or (D)
The compositions required are, in particular, liquid, especially aqueous systems, as mentioned above. (D) above contains a composition as defined above or a composition to be used in conjunction with the enema device as defined above or separately contained in an enema device or similar device. It is intended to encompass any enema means known or commonly used in the art, including kit components.

Suitable enema devices are, for example, known gravity or pump delivery systems, such as rectal and syringe type.

The pharmaceutical composition of this invention is suitably an aqueous solution, dispersion, suspension or emulsion of the selected cyclosporin. Since cyclosporins themselves are highly lipophilic and almost insoluble in water, the compositions of this invention preferably also include (a) a solubilizing agent. Suitable solubilizers include, for example, reaction products of natural or hydrogenated vegetable oils with ethylene oxide, such as those known and commercially available under the tradename Cremophor, such as Cremophor EL [Fiedler, Lexicon der Hilfstoffe]. Lexik
(on der H1lfstoffe), revised and expanded 2nd edition (1981), p. 257 et seq.).

Suitably, the compositions of the invention also include (b) components that increase the overall viscosity of the composition, such as cellulose or cellulose derivatives such as sodium carboxymethyl cellulose, and/or polyols such as sorbitol.

The composition of the invention comprises, for example, dissolving cyclosporin in component (a), for example chromophore, and combining the resulting premix with other additives as mentioned above, for example (b).
Ingredients, such as sodium carboxymethylcellulose/
If sorbitol is present, it is suitable to produce it by dissolving it together with water. To improve the handling properties of the initially produced premix, it is appropriate to include a viscosity reducing agent such as ethanol therein. To avoid precipitation of the cyclosporine component upon standing, the premix can be mixed with the other components immediately before use. Appropriate component ratios are as follows.

(Premix) 1) "Cyclosporin" 2) Cremophor EL 3) Ethanol ratio - Cyclosporin, Cremophor = 1.11-20
, preferably 1:1313-15 (pp ethanol content -0.1-5.0, preferably 0.25
-2.0 (pp based on total amount of 2) (aqueous medium) 4) Water 5) Sodium carboxymethylcellulose 6) Sorbitol ratio - Cyclosporine: (5): (6) = 100
:0.1-5:0.5-1o, preferably 100:0.25-0.5:2.5-5 about 1+20-1+150, preferably about 1=100p
The premix is dissolved in an aqueous medium defined in proportions of pw to obtain a final composition that is stable for a period of several days and is suitable for enema or urethral instillation applications.

The amount of cyclosporine used in practicing the methods of the invention will, of course, vary depending on the particular cyclosporine selected, the condition being treated and the desired effect.

However, - when carrying out the method of the invention for boat fishing, a suitable daily dose is from about 0.5 to about 15.0, preferably about 1
．． 〇 - about io, o, more preferably about 2.0 - about 8.
Coglkg, such as about 3.5 yg/kg or about 7
, on the order of 1 x g/kg, administered once or twice daily by enema.

Each dose is about 17.5 to about 1,000, preferably about 35 to about 700, more preferably about 70 to about 550, such as about 125, about 25019 or 500x
9 cyclosporine.

Suitable cyclosporin concentrations for compositions according to the invention are about 0.5 to about 12.0, preferably about 1°0 to 10.0,
More preferably about 2.0 to about 7°0H/xQz, for example about 2.0 to about 7°0H/xQz. 5y, g/xQ or about the order of 5R9/xQ.

In practicing the invention, contact of the enteral dosage form is suitably prolonged, eg using dosage compositions that remain in the enteral tract for at least 15, eg 30 minutes.

A preferred cyclosporin used in practicing the present invention is "cyclosporin." A more preferred cyclosporin is known as cyclosporin G [N v
a] ”-cyclosporine.

The following examples illustrate the invention in various embodiments.

[Example] Production premix of cyclosporin enema composition ■, cyclosporin, e.g. "Cyclosporin J 250mg2, Cremophor EL"
3.25g 3.94% ethanol
Approximately 834 mg for a final volume of 5 ml.

Aqueous medium 4, sorbitol 5.5g5, carboxymethylcellulose/Na 600mg6, pharmaceutical grade H,O final amount 9Foal.

The premix and aqueous medium are prepared and homogeneous using conventional pharmaceutical techniques. The latter and the former were combined and mixed, with a final volume of 100 ml and a cyclosporins content of 2
A homogeneous composition with 50 mg (= 2, 5 mg/@1). The composition is filled into sterile containers and made into single dosages for use in this invention.

The dose to be treated is provided to the subject along with an enema device consisting of a plastic cylinder with a short plastic catheter. Introduce the dose into the barrel of an enema device for administration by enema.

For example, as a retention enema, each enema for an average 70 kg adult contains 3', 5 mg/kg of cyclosporine.

A similar composition may be prepared containing 500 mg of cyclosporins, such as "cyclosporin" instead of 250 mg of that used above.

"Cyclosporine" in the above composition in similar or equal amounts
Alternatively, an equivalent formulation containing [Nva]'-"cyclosporine" as the active ingredient can be prepared.

[Clinical trial] ■ Treatment of colonic disease with enema (-next trial) The test subjects had ulcerative colitis and chronic active proctitis, which were resistant to topical corticosteroids, and were treated with 5-ASA enema. It consists of the sounds of eight outpatient patients undergoing treatment. Median disease duration was 6 years (range 2 to 22 years).

The target contains "cyclosporine" as component 1,
100ml of the composition of the above example was received. The composition was administered once daily in the evening as an enema and retention enema as described above. Thus, this composition provides a daily dose of 250 mg of "cyclosporine" administered locally in the colon.

Single-dose pharmacokinetic studies were conducted in 7 patients during the initial enema administration period. Venous whole blood samples were taken before and during the following hours overnight. 30 for 4 hours
every minute, every hour for 4 hours, every 2 hours for 8 hours, and then after 24 hours. Another blood sample was taken in the morning two weeks after treatment I. Whole blood "cyclosporine" concentrations were measured by RIA [Sandoz (Polyclonal) kit (Basel, Switzerland)].

Subjects were evaluated 2 weeks after treatment and at 4 weeks follow-up. Blood, pus and mucus passage were each evaluated on an ordinal scale of 1 to 4. 1) None, 2) Less than once a week,
3) at least once a week, and 4) daily. Binder et al., Scandinavian Journal of Gastroenterology (Scand.

J, Ga5troentero1), vol. 5, 627
~632 pages (1970), Gastroenterob (
G astr6entero1ogy), vol.83, 56
The degree of inflammation at sigmoidoscopy activity and rectal examination resection according to the method described by Mon, pp. 3-568 (1982) was also evaluated on an ordinal scale from 1 (normal appearance) to 4.

[Results With the exception of one dose of 9 ng/ml, blood samples taken during the 24-hour follow-up did not show detectable cyclosporin concentrations (<63 ng/ml) and
It was also absent in a blood sample obtained two weeks after treatment I. Repeat sigmoidoscopy showed no signs of mucinous toxicity resulting from the procedure. The only side effect recorded was nausea (
(in 2 patients). No other adverse results were encountered and no changes were observed in plasma creatinine, plasma potassium or liver fraction tests.

Clinical and sigmoidoscopy improvement was seen in 6 patients. Complete remission of symptoms was seen in 21 patients, substantial improvement in 2 patients, and slight improvement in 2 patients. At follow-up, three subjects showed further improvement sigmoidoscopically and clinically. Two subjects were recorded as non-responders. Substantial improvement on histology was seen in 2 subjects (repeat resection was not possible for all subjects). Planned colonostomy was canceled for one subject.

Il, Treatment of diseases of the colon by enema (public study) The subjects consisted of a group of patients exhibiting (i) ulcerative colitis and (ii) Crohn's disease. Each group consists of approximately 6 subjects, male and female, age range 18 to 65 years, presenting with symptomatic and sigmoid rectitis. In the case of subjects exhibiting ulcerative colitis, the selected subjects are [refractory to conventional treatments].
Subjects classified as having Crohn's disease, i.e., exhibiting minimally or moderately active disease despite any prior treatment, or having Crohn's disease activity index I50 despite any prior treatment. There is [gust a
x7 astroenterology
), Vol. 86, pp. 249-266 (1984)]. Subjects in the ulcerative colitis group exhibit normal hardness and softness. The following exclusion criteria were used.

7'IL Systemic steroid treatment within 30 days of ulcerative colitis.

Crohn's disease group: >20 mg/day systemic steroid treatment;
Change in steroid dose within the past 14 days, involvement beyond the rectum.

Both groups: topical steroids within the past 14 days or 5-
Use of ASA therapy, metronidazole therapy, or change to oral rosalazovirine therapy. Any current or past malignancy, S-creatinine > 110 mmol/L nephrotoxic drugs [aminoglycosides, sulfamethizole,
co-treatment with drugs that interact with the absorption of ``cyclosporine'' [phenytoin, palpitate, carbamazepine, INH, erythromycin, rifampicin, ketoconazole, blockers of calcium channels]; Concomitant treatments, damage to liver function [bilirubin, transaminases, alkaline phosphatase, any of these raised to more than twice the upper limit of the normal range], hereditary angioedema, epilepsy, uncontrolled hypertension, ulcers. Reduced life expectancy due to acute or chronic illness unrelated to genital colitis or Crohn's disease, uncontrolled bacterial, viral and/or fungal infections, acute peptic ulcers, non-compliant subjects, pregnant women and childcare. women, alcohol and/or
or substance abuse, very poor dental hygiene.

Using the compositions of the examples as described in item (■) above, the subject was administered once daily before bedtime (daily dose = 250 mglL using enema administration). Subjects were asked to lie down on their left side for 30 minutes. Treatment continued for 2 weeks.

Subjects were evaluated pre-treatment, 1 and 2 weeks post-treatment, and 1 to 4 weeks post-treatment when available for follow-up.

The following tests were performed at each visit.

Sigmoidoscopy. Three test excisions were taken.

One was used for histology and two were immediately sealed, frozen in liquid nitrogen and stored at -80°C for later immunohistochemical studies. Sigmoidoscopy activity and histological expression were quantified as described in Binder et al., supra.

Clinical activity was recorded by completing an examination form. For the ulcerative colitis group, Binder et al.
9-444, (1,976)].

[Blood test] 12-hour trough (Lr) by HPLC or RIA
Measurement of cyclosporine levels in the ulcerative colitis group The following evaluation method was performed for the ulcerative colitis group.

■ Normal intestinal activity once or twice a day without disease inactivation and presence of blood. No signs of underlying disorder.

++, slightly active disease: presence of colonic activity more than 2 times a day, but up to 4 times and/or a small amount of fresh blood. No signs of underlying disorder.

Ill, moderately active disease: colonic activity and/or loose blood in the stool more than 4 times a day. No signs of underlying disorder.

■Very active disease: Same as III except that it is accompanied by severe and essential disorders such as fever, weight loss, anemia, etc.

Sigmoidoscopy activity: 1. Normal: smooth and shiny mucous membrane ++, slightly active: slightly grainy and/or brittle.

Ill, moderate activity 1 marked granularity and friability with diagnostic ulceration.

Histological grade 1, normal Il, slightly active: normal surface and crypt epithelium, but slightly irregular crypt epithelial organization and/or signs of increased cell number in the fixed layer.

Ill, moderate activity: flat surface and crypt epithelium in some areas. Rugged crypt configuration and increased cell number in the fixed layer.

■, a very active biepithelial surface and a completely flat, lacking some areas, a lesser degree of crypt epithelium. Rugged crypt configuration. Obvious inflammatory cell infiltration in the fixed layer.

The following evaluations were performed in the Crohn's disease group.

Types of sigmoidoscopy: fissure, attho
uS) Ulcer, oesema, terrible.

Histological classification, presence of granulomas.

The following blood tests were performed (B-whole blood, P plasma, S=
serum). B-erythrocyte sedimentation rate, B-hemoglobin, B-
hemadcrit, B-white blood cells, B-thrombocytes, P-creatinine, P-potassium, P-bilirubin,
P-A, 5ATSP-ALATSS-albumin, P-
Urate, P-orosomcoside, B-r cyclosporine J
, 12 hour trough.

Absorption of Cyclosporine was measured in 6 subjects. 1
Subjects were admitted to the hospital during the day or in the afternoon of the study. After the enema was performed at approximately 8:00 p.m., blood samples were taken from the venous plastic cannula every hour for the first 12 hours and then every 2 hours for the next 12 hours. 1
0ml whole blood and serum from 5a+1 tubes were collected each time. Whole blood ``cyclosporine'' concentrations were measured by RIA.

The remainder of the blood was deep frozen for later HPLC measurements. Serum was also deep frozen for later immunological studies. Sigmoidoscopy was performed on the second day of admission (afternoon) for safety reasons.

All results obtained in the above studies showed significant improvement in both Crohn's disease and ulcerative colitis groups, with disease control/remission recorded in the majority of subjects and fewer hyporesponses. Ta. A substantial proportion of subjects may be classified as having good or very good disease control/remission of symptoms.

In all cases, the treatment is well tolerated with no untoward side effects other than nausea in a small number of subjects at the beginning of treatment. No side effects were recorded that were harmful to the mind or body, and the number of records of the ``cyclosporine'' serum that was harmful to the mind and body was minimal.

With the above or equivalent treatment methods according to the invention, positive results may be obtained as well using tests using patients with other diseases of the intestine, such as those previously described herein.

The dose of cyclosporine used in studies such as those described above may vary within the ranges previously described herein. As an active ingredient, [N instead of cyclosporine J]
va]” - “cyclosporine” can be used at similar or equivalent concentrations to achieve equivalent results.

Claims (9)

[Claims] (1) A method for treating intestinal disease in a subject in need thereof, comprising locally administering a therapeutically effective amount of cyclosporine into the intestine of the subject. (2) A method of administering cyclosporine therapy for intestinal disease in a subject in need thereof, comprising administering a therapeutically effective amount of cyclosporine by enema. (3) The method according to claim 1, wherein the cyclosporin is "cyclosporin." (4) The method according to claim 1, which is for the treatment of intestinal autoimmune diseases. (5) The method according to claim 4, which is for the treatment of inflammatory bowel disease, Crohn's disease, ulcerative colitis, proctitis, sigmoiditis or complex rectosigmoiditis. (6) Cyclosporine about 0.5 to about 15.0 mg/k
2. The method of claim 1, wherein the method is administered at a daily dose of g. (7) About 2.0 to about 8.0 mg/kg of cyclosporine
7. The method of claim 6, wherein the method is administered in a daily dose. (8) A pharmaceutical composition in the form of an enema containing cyclosporin as an active ingredient. (9) Enema device means containing or delivering a pharmaceutical composition containing cyclosporine as an active ingredient.