AU610561B2 - Utilization of morphine antagonists in the preparation of drugs having an immunomodulator and antiviral effect, particularly for treating acquired immunodeficiency states - Google Patents

Abstract

Utilization of morphine antagonists, such as Naltrexone, beta-endorphine 1-27, or their derivatives or analogs, in the preparation of an immunomodulator and/or antiviral drug for the treatment of acquired immunodeficiency states, particularly the infection by HIV.

Description

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11 AU-Al -11896/88 CT ORGANISATION NINIL D RPR IF *NIV1E QL DEMANDE INTERNATIONALE PUBLIEE EN VERTU DU TRAITE DE COOPERATION EN MATIERE DE BREVETS (PC7) (51) Classification internationale des breyets 4 Num6ro de publication internationale: WO 88/ 05297 A61K 33/4830 /2 7/4A (43) Date de publication internationale:- 28Sjuillet 1988 (28.07.88) (21) Num~ro de la demande Internationiale: PCT/FR88/00032 Publi~e Avec rapport de recherche internationale.

(22) Date de d~p~t international: 21 janvier 1988 (21,01.88) Avant I'expiration dit dci/ai pr~wt pour la mnodification des revendications, sera republihe si de telles nzod iications sont regues.

(31) Num~ro de la demande prioritaire: 87/00670 (32) Date de priorit6: 21 janvier 1987 (21.01.87) (33) Pays de priorit6: FR (71)(72) Diposant et laventeur: SI-ELLY, Marc, Yves, Franck, C16ment [FR/FR]; 1, cour de Rohan, F-75006 Paris (FR).

(74) Mandataire: NONY, Michel; Cabinet Nony Cie, 29, rue Cambac~r~s, F-75008 Paris (FR).

A.O.J.P. 1 5 SEP 1988 (81) Etats d~sian~s: AU, BJ (brevet GAPI), CF (brevet GAPI), CG (brevet GAPI), CMv (brevet GAPI), DK, GA (brevet GAPI), JP, ML (brevet GAPI), MR (bre- ASRLA vet GAPI), SN (brevet GAPI), TD (brevet GAPI), TG ASRLA (brevet GAPI), US. PAU OFF8C (54) Title: UTILIZATION OF MORPHINE ANTAGONISTS IN THE PREPARATION OF DRUGS HAVING AN IMMUNOMODULATOR AND ANTIVIRAL EFFECT, PARTICULARLY FOR TREATING ACQUIRED IMMUNO DEFICIENCY STATES (54) Titre: UTILISATION D'ANTAGONISTES DE LA MORPHINE DANS LA PREPARATION DE MEDICA.- MENTS A EFFET IMMVUNOMODULATEUR ET ANTI-VIRAL, DESTINES NOTAMMENT A TRAI- TER LES ETATS IMMAUNO-DEFICITAIRES ACQUIS (37) Abstract Utilization of morphine antagonists, such as Naltrexone, beta-endorphine 1-27, or their derivatives or analogs, in the preparation of an immunomodulator and/or antiviral drug for the treatment of acquired immunodeficiency states, particularly the infection by H-IV.

(57) Abr~g6 Utilisation d'antagonistes de la morphine, tels que Ia Naltrexone, la b~ta-endorphine 1-27, ou leurs di~riv~s ou analogues, dans Ia preparation d'un mndicament immunomodulateur et/ou antiviral destin6 notarnment d~ traiter les 6tats immunio-d~ficitaires acquis, en particulier l'infection par le VIH.

signature of declarant(s) (no attestationl required) Note: Inital all alterations.

Y

DAVIES COLLISON, IMELBOURNE and CANBERRA.

COMMONWEA4LTH OF AUSTRALL PAIENT ACT 1952 -COWETSPECO~Nt~ NAME ADDRESS OF APPLICANT: Marc Yves Franck Clement Shelly 1, Cour de Rohan Paris 75006 France NAMAE(S) OF INVENTOR(S): Marc Yves Franck Clement SHELLY ADDRESS FOR SERVICE: DAVIES C')LLISON Patent Attornt:.ys 1 Little Collins Street, Melbourne, 3000.

COMPLETE SPECIFICATION FOR THE INVENTION ENTITLED: Utilization of mnorphine antagonists in the preparation of inununomodulator and antiviral effect, particularly for finuno deficiency states The following statement is a full description of this invention, including performing it known to me/us:treating acquired the best method of i r The object of this invention is the use of morphine antagonists in the preparation of drugs producing. an immunomodulator and antiviral effect, intended for treating acquired immunodeficiency states, and in particular, infections caused by the HIV virus (Human Immunodeficiency Virus).

It is well-known that immunodeficiency conditions are characterized especially by a pronounced depression of immunesystem defenses acting through cell intervention. Proposals have already been made to treat these conditions using a wide variety of agents or therapeutic techniques capable of stimulating cellular immunity by means of various mechanisms.

The conventional immunostimulation approach is actually based on the hypothesis that depressed immune-system functions in these pathological states must be activated using general transmitter substances for communication between immunocompetent cells. These transmitting substances, which possess no narrow physiopathological specificity,-prove to be generally unsuitable in this indication, and their undesirable side effects most often catse abandonment of their use.

The observed failures of conventional immunostimulants do not really surprise the specialists, who are acquainted with both the wide variety of immune-system defense mechanisms and the la A Ir complex, numerous, and as-yet unexplained interactions between these various mechanisms. The notion of 'an .immunostimulant actually embraces substances producing very diverse, and indeed even opposing, effects on some categories of immunocompetent cells. For this reason, specialists often prefer to call these substances immunomodulator agents.

Immunologists are well are that the in vitro, or even in vivo study (which, in any event, necessarily remains fragmentary) of the action produced by potential immunomodulating agents on the various immunocompetent cells, prevents them from anticipating the nature of the total effect of these agents on patients already presenting with immune-system dysfunctions.

This situation is, of course, even truer when one considers the treatment of persons suffering from acquired immunodeficiency syndromes caused by infection with the HIV virus, given the many failures already observed in this area.

Furthermore, the use, particularly in regard to HIV infection, of antiviral substances such as viral replication inhibitors, azidothymidine (AZT), is already well-kncwn.

However, because of major toxic effects, the risks associated with this drug, especially for immunodepressed subjects, restrict its use to serious cases of infection which threaten the prognosis for survival (see, for example, Example D, INGRAND, "Medecine et Maladies infectieuses" ("Medecine and Infectious Diseases"), November 1987, Volume 17, N' 11 bis, page 672).

It has now been discovered, surprisingly, that morphine antagonists, which are in particular endowed with an affinity for mu-receptors of opioids, make it possible to styrengthen the immune-system, cell-mediating defenses of patients suffering from acquired immunodeficiency states, and especially of subjects infected with HIV. These antagonists possess, furthermore, an intrinsic antiviral action, especially against the HIV.

The fortuitous observation of a marked clinical improvement in a number of HIV-infected subjects, former drug addicts undergoing treatment with Naltrexone as a support treatment following withdrawal, led the applicant, first, to study more in depth the action of this drug on the strengthening of immunesystem, cell-mediated defenses and its possible antiviral action, and second, to find out whether the same beneficial effects could be observed using other morphine antagonists.

It is well known that morphine and some opioids possess a strong linking affinity for mu-receptors, and secondarily for the kappa-receptors found on some nerve cells. It is also known that mu-receptors are also found on some immunocompetent cells. Other endogenous opioids, especially.- Met-enkephalin, activate the socalled delta-receptors, which, once again, are also found both on nerve cells and on some immunocompetent cells.

It is now known that the activation of opioid receptors (currently called mu-receptors) by morphine and some exogenous opiates produces an immunodepressive effect, and that the 3 O ~'7 Cji~ activation of other receptors (currently termed delta-receptors), especially by Met-enkephalin, produces an. immunostimulating effect (see, for example, WYBRAN et al., Journal of Immunology, 123(3), 1068, 1979).

Although the applicant does not consider himself bound by this hypothesis, the immune-system stimulation effect and the antiviral actions observed clinically with Naltrexone can be explained schematically by supposing that opiate-containing factors (alkaloids) and/or opioids (peptides) not yet identified produce an immunodepressive, infection-promoting effect mediated by the activation of the morphine-like mu-receptors and, secondarily, of the kappa-receptors, and that Naltrexone is antagonistic to this dual effect.

To verify the validity of this hypothesis, the applicant studied the effect of beta-endorphin fragment 1-27. It is known that beta-endorphin, a peptide composed of 31 units of amino acids, has the capacity to activate the mu- and delta-receptors simultaneously, the. morphinomimetic effect linked to the stimulation of the mu-receptor predominating, however (see Example LOH et al., Proceedings of the National Academy of Sciences (USA), 73, 2895 (1976).

It is also known that beta-endorphin fragment 1-27, which is one of the products of degradation of beta-endorphin in the organism, possesses an effect antagonistic to the beta-endorphin effect as regards the mu-receptor (see, for example, HAMMONDS et al., Proceedings of the National Academy of Sciences (USA), 81, 1389 (1984).

Now, the applicant observed that beta-endorphin fragment 1- 27, like Naltrexone, makes it possible to strengthen the cellmediated, immune-system defenses while at the same time producing an intrinsic antiviral effect. This observation is therefore compatible with the hypothesis put forth above.

Other, possibly complementary hypotheses may be formulated.

For example, the occupation of mu sites by Naltrexone or fragment 1-27 may promote the occupation and activation of delta-receptors by beta-endorphin.

It is also possible that Naltrexone -otects and even promotes the release of endogenous enkephalins by attaching itself to the presynaptic autoreceptors capable of inhibiting this release.

An intrinsic antiviral effect has also been observed in cases of HIV infection.

Indeed, in the patients mentioned above who underwent treatment with Naltrexone, a noticeable improvement in the following clinical symptoms, was observed: fever, general discomfort, and night sweats, as well as restoration of appetite and weight gain. It is known that these symptoms, whose disappearance or improvement is observed here, typify viral replication (see, for example, R. Yarchaon et al., in AIDS: Modern Concepts and Therapeutic Challenges, S. Broder, ed., L L I c Marcel Dekker, Inc., p. 343 (1987).

The antiviral effect observed in cases, of HIV infection using Naltrexone may be explained schematically, for example, by forming the hypothesis that some viral proteins in acquired immunodeficiency states behave as do morphinomimetic substances, either directly or through the mediation of endogenous substrates.

It should be noted that, in contradistinction to conventional treatments, the invention treatment using morphine antagonists amounts to the production, first, of an immunomodulator effect, not by stimulating generally immunesystem defenses, but by inhibiting an immunosuppressive effect specific to the disease, and second, an antiviral activity. In this particular case, the therapeutic distinction between these two immunomodulator and antiviral effects may no longer be drawn, since the morphine antagonists possess a dual pharmacological capability.

The totality of the observations made regarding the effects of Naltrexone and of beta-endorphin fragment 1-27 makes it possible, therefore, to recommend the use of morphine antagonists in the treatment of acquired immunodeficiency states, and, in particular, of HIV infection.

The object of the present invention consists, therefore, of morphine antagonists, especially in their interactions with the mu-receptor, like the active ingredients in the preparation of an 6

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\q L i_ _I I;immunomodulator and/or antiviral drug intended for the treatment of acquired immunodeficiency states, and 'especially of HIV infections.

Among the morphine antagonists which may be used according to the invention, Naltrexone and its structural counterparts (such as beta-Chlornaltrexamine, Nalmephene, Naloxazone, Naltrexonazine, etc., as well as their derivatives, including acceptable pharmaceutical salts, and especially their chlorhydrates, citrates, sulfates, etc., and the corresponding Nquaternary derivatives) may be mentioned. N-quaternary derivatives are, for example, quaternized by means of alkylation of these compounds with a lower alkyl group containing, for example, from 1 to 4 carbon atoms. These derivatives are conventionally prepared. Naltrexone is currently preferred, because its safety in use has already been demonstrated during the support treatment of opiate-dependent drug addicts following the withdrawal period, as recommended in US Patent N" 3.332 950.

The long-Lerm use of Naltrexone actually produces no major undesirable effects, and is characterized by the absence of habit-forming conditions and- by a high tolerance to the recommended dosages. A further advantage of Naltrexone is its ability to be administered orally and to easily penetrate the blood-brain barrier.

In some cases, preference may be given to the use of strictly peripherally-acting drugs which do not penetrate the I blood-brain barrier, as is the case with N-quaternized derivatives of Naltrexone and its counterparts, for example a halide such as the N-methyl Naltrexone bromide.

It will be recalled that Naltrexone is the 17- (cyclopropylmethyl) -4,5-epoxy-3,14-dihydroximorphinane-6-one..

Among the morphine antagonists which may be used according to the invention, we may also mention the beta-endorphin fragment 1-27, and, more generally, peptides containing from 6 to 30 units of amino acids and whose C-terminal end incorporates the minimum sequence of formula I or II: Ile-Ile-Lys-Asn-Ala-His-CO 2 H (I) Ile-Ile-Lys-Asn-Ala-Try-CO 2 H (II) which is responsible for attachment to the mu-receptors. We may also include the derivatives and analogues of these peptides containing modifications intended, for example, to avoid "heir rapid degradation by peptidases or to promote their penetration through the blood-brain barrier.

These peptides are prepared conventionally, either by the degradation of natural peptides or by synthesis or hemisynthesis.

similarly, the preparation of derivatives of these peptides (not easily degradable), which involves, for example, insertions of Nmethylated or configuration D amino acids, or of other synthetic amino acid derivatives, is carried out using well-known 8 _1 i. procedures (see, for example, B. Rajashekhar et al., Journal of Biological Chemistry; 261 13617 (1986).

Hammonds et al, in the article cited, have described the antagonistic action of morphine for beta-endorphin 1-27.

Morphine antagonists also include deacetylated alphamelanotropine (also called alpha-MSH); or some ACTH fragments not producing corticotropic activity, such as fragments 1-16, 5-16, 5-14 and D-Phe-ACTH 4-10 ("Phe" represents phenylalanine) (see Gispen, European Journal of Pharmacology, 39.393-397 (1976).

The preparation of drugs containing a morphine antagonist is carried out in a conventional manner. For example, the active ingredient is mixed with a suitable pharmaceutical diluting agent, vehicle, admixture, or excipient allowing oral, rectal, vaginal, or parenteral (including subcutaneous, intravenous, endonasal or intrathecal) administration of the drug.

The proportion of the active ingredient present in these drugs is normally 0.05 to 50% by weight.

Oral administration is generally reserved for non-peptidecontaining ingredients, and endonasal administration, for peptide-containing ingredients. Intrathecal administration may be used for both types of active ingredients, and may require, if necessary, a constant- or variable-flow, continuous or discontinuous ad hoc perfusion apparatus.

The pharmaceutical embodiments which may be used are, in particular, the following: coated or uncoated tablets, pills, capsules, divisible tablets, tablets with a protective coating, time-release tablets, microcapsules having an enteric coating, soft capsules, capsules, pills, cachets, injectable aqueous solutions, solutions used as nasal sprays, suppositories, capsules for gynecological use, and drinkable solutions, syrups, or suspensions, or freeze-dried preparations that can be reconstituted at the time of use to make up injectable solutions.

Dosages prescribed for drugs producing an immunomodulator and/or anitviral effect according to the invention vary based on the mode of administration, the frequency of treatment, the phase of the disease, the age and weight of the patient, and the particular reaction of the patient.

In general, from 0.02 to 2 mg of active ingredient are administered per kg of body weight per day.

As regards Naltrexone and its derivatives and structural analogues, the daily average oral dose is normally 30 to 70 mg (and especially 50 mg) of active ingredient per day for an adult, taken once or several times. The dosage is preferably increased gradually. For example, the oral dose is from 5 to 10 mg the first day, 20 mg the second day,, from 25 to 30 mg the third day, etc., up to the desired daily dosage. In cases of drug-addiction to exogenous opiates, these drugs will preferably be administered only after the patient has has ceased using the opiate for 7-10 days.

i0 f In the cases of patients who react only minimally to these drugs, higher doses may be administered for, a limited period, doses of from 150 to 500 mg of active ingredient per day for an adult.

When using peptide-containing derivatives as active ingredients according to the invention, from 0.02 to 0.1 mg per kg per day for an adult is generally administered parenterally, especially subcutaneously, endonasally, or intrathecally.

Reduced dosages are used for intrathecal administration; 0.05 to 0.1 mg/kg of active ingredient per day, whether peptide-containing or non-peptide-containing.

The drugs described above may be administered, for example, to subjects suffering from acquired immune deficiencies, especially in cases of HIV infection, from immunodepressive states, including those associated with pregnancy, and from diseases having an auto-immune component. Administration to pregnant women is not counterindicated, since the active ingredients used produce no teratogenic effect.

Drugs according to the invention may be used in the treatment of infections caused by opportunistic viruses (for example, herpes, Epstein-Barr, cytomegalovirus, papillomavirus, etc.).

An initial 1-3-month treatment period is normally recommended. This period may be extended based on clinical assessment. During treatment with Naltrexone or its analogues I I -I I 1 and derivatives, the regular monitoring of serum transaminases is recommended to monitor hepatic tolerance.

These drugs producing an immunomodulator and/or antiviral effect may, in accordance with the invention, be used in combination with other antiviral or antimitotic chemotherapies, as required.

They may also be used in combination with substances that potentiate the action of endogenous opioids producing an immunostimulating effect, such as Met-enkephalin, a ligand of delta-receptors. These potentiating substances include zinc (in the form of acetate, orotate, or any other acceptable pharmaceutical salt), at a dosage, for example,, of 20 to 50 mg per day for an adult, taken orally), or enkephalinase inhibitors.

They may also be associated with aminopeptidase inhibitors, with beta-endorphin-release inhibitors such as dopamine, L-dopa, bromocriptin (dopaminergic antagonist), or peripheral and central receptor antagonists of benzodiazepines, especially triazolated derivatives.

Oral compounds containing zinc will also preferably contain a sufficient quantity of lactose, for example 75-90 my per unit dose, to promote the digestive absorption of the zinc.

Certain nutriments promoting the immune response, such as selenium, vitamins E and C, the B vitamins, especially B6 and B12, folates, or certain polyunsatured fatty acids may, furthermore, be incorporated into drugs producing the 12 I 4VS.

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immunomodulator and antiviral effect according to the invention.

Another object of the invention, as regards. immunomodulator and/or anitviral drugs, includes peptides selected from among those containing from 6 to 30 amino acids, whose C-terrainal end has the minimum sequence of formula I or II (given above), or the deacetylated alpha-MSH and ACTH fragments mentioned above, as well as the analogues or derivatives of these peptides. These drugs contain the active ingredient in combination with a suitable pharmaceutical vehicle, excipient or diluting agent, in addition to the active ingredient present, for example, in the proportion of from 0.05 to 50% by weight.

A further object of the invention is a method of treatment of acquired immune-deficiency states, especially HIV infection.

This treatment is characterized by the fact that the patient receives an effective quantity of at least one immunomodulator and/or antiviral drug, such as one described above.

The following examples illustrate the invention without restricting its scope.

Examples of Pharmaceutical Compounds In a preparation intended for oral administration, such as a tablet, the active ingredient, such as Naltrexone, for example in a 50-mg dose per administered unit, is combined with lactose and

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4 an inert, non-toxic, pharmaceutically acceptable vehicle given by mouth, such as pharmaceutical quality starch.

Suitable binding agents, lubricants, decomposition and coloring agents may also be incorporated.

Non-restrictive examples of bonding agents include starch, gelatin, cellulose, lactose, and waxes.

A non-restrictive example of a lubricant to be used in the dose-measured forms is stearic acid.

Suitable decomposition agents include, for example, starch and cellulose.

If desired, a pharmaceutically acceptable, conventional coloring agent may also be added, such as azorubine, quinoline yellow, or indigotin.

Finally, a coating or varnishing may be provided.

The single tablets may contain solely the active ingredient mentioned above, combined or mixed with a pharmaceuticallyacceptable, inert, non-toxic vehicle or excipient; administration of dosages may also be in the form of so-called biphasic tablets, in which a suitable coating such as a natural polymer like gelatin, or a fatty glycerol ester such as precirol, separate, while maintaining stability, the active ingredient from the zinc acetate and the lactose, after the varnishing of this first core; finally, there are so-called "three-layer" tablets produced using conventional industry techniques.

14 r Parenteral administration requires the preparation of isotonic aqueous solutions, possibly with a preservative.

Freeze-dried compounds based on aqueous solutions, generally containing a lyophilization additive such as mannitol, lactose, or similar substance, may be prepared. These lyophilized compounds are reconstituted when administered as injectable or sprayable solutions. Injectable solutions in the form of lyophilized solutions or powders may contain a zinc salt.

EXAMPLE 1 Tablets having the following composition are prepared conventionally: M_/tablet Naltrexone (chlorhydrate)...... lactose sodium 2 stearic acid One tablet is given daily. Tablets containing measured 7 EXAMPLE 1 sodium 2 'V 0 10 i

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EXAMPLE 2 So-called biphasic tablets are made according to standard methods; they are composed of an initial core separating, while maintaining stability, the active ingredient from the zinc acetate and the lactose by means of a dry coating formed using precirol as a binding agent, followed by suitable varnishing.

Naltrexone (chlorhydrate).......

zinc starch magnesium stearate stearic mcg/tablet 4 2 One tablet per day is administered to adults.

EXAMPLE 3 Using standard methods, so-called biphasic tablets are made in which a first core separates, while maintaining stability, the active ingredient from the zinc acetate and the lactose, by means of a dry coating formed using gelatin, followed by a suitable varnishing: 16 ii

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a Kq/tablet Naltrexone (chJlorhydrate) zinc a4cetate lact ose... magnesium stearic 2 Adults take one tablet daily.

EXAMPLE 4: INJECTABLE SOLUTION The composition of this solution for 1 cm 3 is as follows: Naltrexone 0.5 Sodium *9 Methyl parabenzoate 5 Propyl parabenzoate 0.1 Apyrogenic distilled water, gsp 1 This solution is divided into ampoules cm 3 ou 5 cm 3 mg Mg mg mg containing either 2 EXAMPLE 5: LYOPH-ILIZED COMPOSITION This composition contains: -Beta-endorphin 1-27 0.3 mg -lactose 30 7mg An injectable soluti=~ is obtained by dissolving this mixture in 3 cm 3 of a sterile, apyrogenic aqueous solution containing 0.9% NaCi.

EXAMPLE 6: AOUEOUS SOLUTION FOR NASAL SPRAYS Composition: B feta-endorphin 30 mg -Excipient (anhydrous disodiun phosphate, sodium chloride, sorbitol, glycerol, preservatives, distilled water) gsp 10 ml.

Approximately 0.1 ml is sprayed in each nostril 4 to 6 times daily.

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Claims (12)

1. Use of morphine antagonists possessing an affinity for mu-receptors or for mu- and kappa-receptors, as active f- ingredients in the preparation of an immunomodulator and/or antiviral drug for treatment of acquired cellular-immunity deficiencies.

2. Use according to claim 1 in the preparation of a drug for treatment of HIV infections.

3. Use according to either of the preceding claims, wherein said antagonist is chosen from among Naltrexone, beta- chlornaltrexamine, Nalmephene, Naltrexonazine, and Naloxazone, as well as their pharmaceutically-acceptable salts and corresponding N-quaternary derivatives.

4. Use according to claim 3, wherein the N-quaternary derivatives are quaternized with a alkyl group. Use according to either of claims 1 and 2, wherein said antagonist is a peptide containing 6 to 30 amino acid units and whose C-terminal end contains the minimum sequence of formula I or II: Ile-Ile-Lys-Asn-Ala-His-CO 2 H (1) Ile-Ile-Lys-Asn-Ala-Tyr-CO 2 H (II), I I. 1 N-methylated or configuration D amino acid analogues of these peptides.

6. Use according to claim 5, wherein said peptide is beta-endorphin fragment 1-27.

7. Use according to either of claims 1 and 2, wherein said antagonist is deacetylated alpha-melanotropine.

8. Use according to either of claims 1 or 2, where said ACTH antagonist is selected from among ACTH fragments, S• 1-16, 5-16, 5-14, and D-Phe-ACTH 4-10. 0o

9. Use according to any of the preceding claims, wherein said drug is intended for administration in association with a pharmaceutically-acceptable zinc salt. Method of treatment of acquired cellular-immunity deficiencies, wherein a patient is given an effective quantity of an immunomodulator and/or antiviral drug as described in any of claims 1 through 9. O

11. Method of treatment according to claim 10, wherein said patient suffers from an HIV infection.

12. A method of treatment according to claim 10 or a use according to claim 1, substantially as hereinbefore described with reference to the Examples. DATED this 26th day of February, 1991. MARC YVES FRANCK CLEMENT SHELLY By His Patent Attorneys DAVIES COLLISON S 1 910227,immdatL82,a:\11896she.res,20 1 UV ;4~s I i I rmanila T, .I INTERNATIONAL SEARCH REPORT International Application No PCT/FR 88/00032 I. CLASSIFICATION OF SUBJECT MATTER (if several classification symbols apply, Indicate all) 6 According to International Patent Classificatlon (IPC) or to both National Classification and IPC Int.Cl. A61K 31/485; A61K 37/02; A61K 37/24; A61K 33/30 II. FIELDS SEARCHED Minimum Documentation Searched 7 Classification System I Classification Symbols 4 Int.Cl. A61K 31/00; A61K 37/00 Documentation Searched other than Minimum Documentation to the Extent that such Documents are Included in thr Fields Searched a III. DOCUMENTS CONSIDERED TO BE RELEVANT 9 Category Citation of Document, 11 with indication, where appropriate, of the relevant passages i2 Relevant to Claim No, Y Pharmacology, vol. 14, 1976 1-4,9 W.F. Geber et al.: "Action of naloxone on the interferon-lowering activity of morphine in the mouse", pages 322- 329, see abstract, table 1 Y Adv. Alcohol. Subst. Abuse, vol. 1, 1982, 1-4,9 No. 3-4 (The Haworth Press, Inc.), A. Falek et al.: "Opiates as modulators of genetic damage and immunocompetence", pages 5-20, see abstract; page 12; page 14, lines 1,2; figure 2 Y Otolaryngol. Head Neck Surg., vol. 94, 1986, 1,2,5-9 G.T. Wolf et al.: "Beta endorphin enhances in vitro lymphokine production in patients with squamous carcinoma of the head and neck", pages 224-229, see pages 227-229; abstract Y Trends Neurosci, vol. 717, July 1984, 1-9 SSpecial categories of cited documents: 10 later document published after the International filing date document defining the eneral state of the art which is not or priority date and not In conflict with the application but A o iredtobe oft errlesane art whcch s nocited to understand the principle or theory underlying the considered to be of particular relevance invention earlier document but published on or after the International document of particular relevance; the claimed invention filing date cannot be considered novel or cannot be considered to document which ihay throw doubts on priority claim(s) or Involve an inventive step which is cited to establish the publication date of another document of particular relevance' the claimed Invention citation or other special reason (as specified) cannot be considered to Involve an inventive step when the document referring to an oral disclosure, use, exhibition or document is combined with one or more other such docu- other means ments, such combination being obvious to a person skilled document published prior to the international filing date but In the art. later tnan the priority date claimed document member of the same patent family IV. CERTIFICATION Date of the Actual Completion of the international Search Date of Mailing of this International Search Report 19 April 1988 (19.04.88) 29 June 1988 (29.06.88) International Searching Authority Signature of Authorized Officer EUROPEAN PATENT OFFICE Form PCT/ISA/210 (second sheet) (January 1985) r Internatlonal Appllc.,on No. PCT/FR 88/00032 2- FURTHER INFORMATION CONTINUED FROM THE SECOND SHEET Chang: "Opioid peptides have actions on the immune system", pages

234-235, see the whole document A Life Sciences, vol. 35, No. 1, 1984 1-9 (Pergamon Press, US), N.Kay et al.: "Endorphins stimulate normal human peripheral blood lymphocyte natural killer activity", pages 53-59, see abstract; page 53, lines 8-16; pages 58-59 (discussion) A The Journal of Immunology, vol. 136, 1,2,5--9 V.Q OBSERVATIONS WHERE CERTAIN CLAIMS WERE FOUND UNSEARCHABLE This International search report has not been established In respect of certain claims under Article 17(2) for the followlng reasons: 1.7 Claim numbers..XX..., because they relate to subject matter not required to be searched by this Authority, namely: xx Claims 10-11 See PCT Rule 39.l(iv) Methods for treatment of the human or animal body by surgery or therapy, as well as diagnostic methods. 2. Claim numbers...5.-9, because they relate to parts of the international application that do not campty with the prescribed require- ments to such an extent that no meaningful international search can be carried out, specifically: The expression "analogues" of certain compounds is not sufficiently clear to define a chemical compound. It is impossible to characterize compounds by their biological activity or by their lack thereof. Claim because they are dependent claims and are not drafted In accordance with the second and third sentences of PCT Rule vI.] OBSERVATIONS WHERE UNITY OF INVENTION IS LACKING This International Searching Authority found multiple inventions In this international application as follows: 1.Q As all required additional search fees were timely paid by the applicant, this International search report covers all searchable claims of the international application. S2.- As only some of the required additional search fees wers timely paid by the applicant, this international search report covers only those claims of the International application for which fees were paid, specifically claims: 3. No required addltional search fees were timely paid by the applicant. Consequently, this international search report is restricted to the Invention first mentioned In the claims: it Is covered by clahn numbers: As all searchableclaims could be searched without effort justifying an additional fee, the International Searching Authority did not invite payment of any additional fee. Remark on Protest SThe additional search fees were accompanied by applicant's protest SNo protest accompanied the payment of additional search fees. Form PCTIISA/210 (supplemental sheet (January 1985) 0 i I Lw" mearme PCT/FR 88/00032 III. ooCUMrUS COmUADIREo TO RELIVANT (CoMNT114M OM TX, uCOMO SNMU Cawory Cae orf wommn. -h l UW1o w rapproormw *f w nM pons I ft..l to C1nw No No. 3, 1 February 1986, (The American Association of Immunologists, US), R.N. Mandler et al.: "Beta-endorphin augments the cytolytic activity and interferon production of natural killer cells", pages 934-939, see abstract; page 934 A Psychother. Psychosom., vol. 42, 1984, 1,2,5-9 Karger AG, Basel, CH), E.G. Fischer et al.: "Beta-endorphin modulates immune functions", pages 195-204, see pages 198,199 ("Interferon") A Cancer Research, vol. 41, April 1981, 1,2,5-9 F. Legros et al.: "Alpha-melanocyte- stimulating hormone binding and biological activity in a human melanoma cell line", pages 1539-1544, see abstract; page 1543, right-hand column, lines 4-16 Y The Journal of Immunology, vol. 132, No. 1, 1,2,5-9 January 1984 (The American Association of Immunologists, US), H.M. Johnson et al.: "Regulation of lymphokine (gamma- interferon) production by corticotro- pin", pages 246-250, see abstract PX Dialog Information Services, file 155: 1-9 Medline 66-88/May, accession number: 06408708 B.D. Purdy et al.: "Current concepts in clinical therapeutics: immunologic treatment of human immunodeficiency virus infections" Clin Pharm (United States) Nov 1987, 6 (11) 6851-65 Form PCT ISAM8 (sum one*() CJSwy 1wS) -i A v I I 4' RAPPORT DE RECHERCHE INTERNATIONALE Demandeinternationaie N PCT /FR 338/0003 2 1. CLASSEMENT OE L'INVENTION (si olusieurs symooles des classification sont sooicoles, leS ifloquer tous) Solon Ia classificaionl internelonale des brevets (CIS) ou A Ia fogs melon Ia classification nationalset atI CIS CIB 4:A 61 K 31/435; A 61. K 37/02; A 61 K 37/24; A 61 K 33/30 If. DOMAINES SUR LESQUELS LA gHCM APORTE flotumentation minimale consulfee Syst~me dle classification Symooies oc classification CIB 4 A 61 K 31/00; A 61 K 37/00 Documentation consult~e autre que la documentation minimale pans la mesure o6i de fels documents font sarlie des domagnes sur lesquels la recherche a poi6 Ill. DOCUMENTS CONSID11RI1S COMME PERTIMENTS, '0 Cat~gor Identification des documents citds,1 avec indication, si necessaire. N, des revendications a grgedes passages pertinents 12 6me5 3 Y Pharmacology, vol. 14, 1976 1-4,9 W.F. Geber et al.: "Action of naloxone on the interferon-lowering activity of morphine in the mouse', pages 322- 329, voir abr~g6, tableau 1 Y Adv. Alcohol Subst. Abuse, vol. 1, 1982, 1-4,9 no. 3-4 (The Haworth Press, Inc.), A. Falek et al.: "Opiates as modulators of genetic damage and imrunocompetence',. pages 5-20, voir abr~g6; page 12; page 14, lignes 1,2; figure 2 Y IOtolaryngol. Head Neck Surg., vol. 94, 1,2,5-9 1986, G.T. Wolf et al.: "Beta endorphin enhances in vitro lymphokine production in patients with squamous carcinoma of the head and neck", pages 224-229, voir pages 227-229; abr~g6 Y Trends Neurosci, vol. 717, juillet 1934, 1-9 Cat~gories sp~ciales de documents cits: aT e document ult~rteur Publi6 ootriaurement Ala date de d~odt it Av doumet d~inisantI'dtt g~~ra cleIs tchnquenonintearnational ou A Ia date de orgorgte et niappartenent Pas cosi r# aom douet psanticlt t erd tieni e non1a! dI A tal techngque pertgnent, mais cite pour comprendre consd~r coma prticli~emen petinetIl Principe ou Ia tlt~orie constgtuant la base de ligyvention E document antdrieur, maim oubli6 A Ia date de d~p6t interna- a X a document Parficuliirement oertgnent: llInviintion revendi- tional ou aprils cette date qu~e ne pout ktre consid~r~e comme nouvelle ou comme itLe* document pouvant jeter un doufe sur une rev~ndicatior, de implittuant une activgf4 inventgve prioritd ou cgt6 pour determine, I& date as publgcatgon d'une a cY a document particuli~rement Pertinent; l'invention raven- amutre citatgon ou pour une ragaun so~cgale (telle qu~iridiclude) dgqu~e ne pout ktre considr~e comme impliquent une aK 0 v document se r4firant A uns divulgation orale, A us usage, A activit6 inentive lorsque Is document est ssaocI6 A un ou une expostgofn ou toum eutres moyons pfusieurs autres documents de m~ine nature, cette cambg- a P m document Publi,6 avant Ia date de d~odf international, maim sermon Otant Ovidente pour una personne du metier. postdrieurement A Ia date d6i priorit6 aveencliquee ai m document qui felt partge de Ia m~me famifle dle brevets IV. CERTIFICATION Date A lequelle Ia recherche internationale a Wf eliectivement Date d'exo~dition Olu pr~sent rapport de recherche iternationale acftevie 19 avril 1983 2 29 J UN 1988 Administration cherg4e de Ia recherchte internationala 0 ti OFFICE EUROPEEN DES BREVETS 'a aatrs NDER PUTTEN Formutaire PCTIISA/2IO (cdauxime fauilt.i) (Janeis 19s5) f- Demands irnternationale N* PCT FR 33 /000 2 A SUITE DES RENSEIGNEMENTS INOIQUCS SUR LA DEUX19ME FEUILLE Chanig: "O0pioid peptides have actions on the immune system", pages 234-235, voir le document en entier A Life Sciences, vol. 35, no. 1, 1984 1-9 (Pergamon Press, US), N. Kay et al.: "Endorphins stimulate normal human peripheral blood lymphocyte natural killer activity", pages 53-59, voir abrr~g6; page 53, lignes 8-16; pages 58-59 (discussion)i A The Journal of Immunology, vol. 136, 1,2,5-9 no. 3, ler f~vrier 1986, V. OBSERVATIONS LORSOUILl A ItTll ESTIMt QUE CERTAINES REVENDICATIONS HE POUVAIENT PAS FAIRE L'OBJET DUNE RECHERCHE, Solon Iarticla 17.2) a) certaines revendications Wont pas fait I'objet d'une recherche Pour le% motifs suivants: 1. F Les revendicationa nubo.,; s rapportent A un obiet A l'dgard duquel as Prisento administration nia pas l'obligation de pro- cAdcn a is rocnrfcni. a savoir: Revendications 10-11 Voir PCT Ri~gle 39.l(iv) M~thodes de traitement du corps humain ou animal par la chirurgie ou la th~rapie, ainsi que m~thodes de diagnostic. 2. MLea revendications numiro:n 5- 9...so rapporient h des parties de I& demands intornationale qul no reopilsant pas lea conditions preacrites dane une moaure telle qu'une recherche significative ne pout Wer afiectuio, pricia~ment: L'expression "analogues" de certains composes n'est pas suffiserent claire poour definir un compos6 chimiaue. C'est impossible de charact6riser des composes par leur activit6 biologique, ou par le manque d'une telle activit6. 3. E Los rovendications nuros.scont des revencllcations d~peridantes at rno aont pas rdVio coritornefi a It d*uewn4 at A In troisAme phesan~ Is riglo 6.4.a) du PT Vt. OBSERVATIONS LORSOU'Ll Y A ABSENCE D'UNITt DE LINVENTION L'ad mini stralo n charg~e de Ia rocherche Internationale a trouvik plusiours Inventions dana Ia prdsente demande Internationale, c'est-A-dire: 1. Comm* toutea lea taaes additlonnolles demandles ont 6t6 pay~es dons lee d~lait. Is pr~sent rapport do recherche internationale ecuvra tout.: le$ rovendicationa do Ia demands Internationale pouvant fairo lobjet d'une recherche. 2.E Corrimo souleerent unt partle des taxes addltionnelos domanc~st a 6t4 payid danas "aia. W. present rapport do rectworti ntarnationale ouvyq aseom4l ccil40 dos rovendicatioIna do I& domando pour Wtaiue4wws Ina taxes ont iti pay*.a, c'eat--drs lea reeticins 3. Aucuno taxo additlonnolte domand~e no Al payde dane lee dilais par Is d~poant. En cone~quence, Ie prdaent rapport do recherche Internationale eat limits A l'Invention mentionn~e an premier dans lea revendicationa; site eat couverte par lea revendications numilros: 4.1 Etant donn*aouo toutaa lea rovondicationa ausceptibloa do faire l'oblet d'une recherche Io pouvaient tas effort particulier juatifiant uns taxo additionnoill, 'ad ministration cnarg~o oe Ia recie rce international. n'a acilicit6 Is paiomont d'aucune taco additionnolle. Remarqu* Quaint h I& r~aerve LI Les taxes addltionnollo, do rochorche 6talent accompagnilea d'une rkagre. du dilposant. Zj Aucune risorvt na mt6 fait@ bora du palemont dea taxes additlonnel~a do recheorche. Formulate. PCTIISA1210 (fouille supplillmentairo (Janvw 1985) -3- Demand* International* 1N PCT/FR 88/00032 (SUITE DES RENSEIGNEMENTS INDIQU49 SUR LA Ill. DOCUMENTS CONSIDtIUS COMME PERTINENTS OEUXIEME FEUILLE) ksenSI~tdwJthon o" docurronts citi. aysG n4.czbr, wjnelce xair.dervw~-~ Ca *oi de gassag. penwinents r~~dctin (The American Association of Immunologists, US), R.N. Mandler et al.: "Beta-endorphin augments the cytolytic activity and interferon production of natural killer cells", pages 934-939, voir abr~gd; page 934 A Psychother. Psychosom., vol. 42, 1984, 1,2,5-9 Karger AG, Easel, CH), E.G. Fischer et al.: "Beta-endorphin modulates immune functions", pages 195-204, voir pages 198,199 ("Inter- feron") A Cancer Research, vol. 41, avril 1931, 1,2,5-9 F. Legros et al.: "Alpha-melanocyte- stimulating hormone binding and biological activity in a human melanoma cell line"f, pages 1539-1544, voir abr~g6; page 1543, colonne de droite, lignes 4-16 Y The Journal of Immunology, vol. 132, 1,2,5-9 no. 1, janvier 1984 (The American Association of Immunologists, US), H.M. Johnson et al.: "Regulation of lymphokine (gamma-interferon) production by corticotropin", pages

246-250, voir abr6g6 P,X Dialog Information Services, dossier 155: 1-9 Medline 66-88/mai, nurn~ro d'accession: 06408708 B.D. Purdy et al.: "Current concepts in clinical therapeutics: immunologic treatment of human immunodeficiency virus infections" Clin Pharm (United States) Nov 1987, 6 (11) 6851-65 Formulair. PCTIISAM2O (fouille addmtonnolie) (janvw