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    1O 38 Abstract The present invention relates to a novel pharmaoeutical composition comprising oxytocinand/or one or more fragment(s) and/or varlant(s) thereof and at least one non-ioniccellulose ether, such as hydroxypropylmethylcellulose, said pharmaoeutical compositionhaving a low pH. The present pharmaoeutical composition has been shown to provide anexceptionally suitable environment for oxytocin, as the stability thereof has increasedsignificantly as compared to previous compositions with this molecule. Thepharmaoeutical composition according to the invention can be used for medical purposes, such as in the treatment of climacteric disorders.
    公开号:SE536091C2
    申请号:SE1150324
    申请日:2011-04-14
    公开日:2013-04-30
    发明作者:KERSTIN UVNäS-MOBERG；Anders Carlsson
    申请人:Pep Tonic Medical Ab；
    IPC主号:
    专利说明:

    536 091 PS54264EN00 2 stress states during the fetal stage. Such conditions can be cardiovascular diseases, such as stroke, heart attack, hypertension and diabetes. In the human body, oxytocin is produced in the paraventricular nucleus, PVN, and the supraoptic nucleus, SON, in the hypothalamus. It differs by only two amino acids from vasopressin, which is also produced in these nuclei. The magnocellular oxytocinergic neurons in SON and PVN send oxons to the posterior pituitary gland from which oxytocin is released into the circulation.
    Parvocellular neurons that originate in PVN extend into områden your areas within the CNS. The oxytocin-producing cells are supplied by nerves through cholinergic, catecholaminergic, seronergic, as well as peptidergic neurons. The presence of oxytocin in various tissues outside the brain, such as the uterus, ovaries, testicles, thymus, adrenal medulla and pancreas, has been detected and oxytocin is released to exert local effects in these organs. A parallel secretion of oxytocin into brain regions and into the circulation occurs in response to certain stimuli such as breastfeeding but other stimuli can cause separate activation of oxytocinergic neurons, which end in the brain or pituitary gland.
    Cellulose ethers are named after, and based on, cellulose, which is a renewable material and is the most common chemical compound in the organic sphere. There is a wide variety of cellulose ethers available on the market, both ionic and non-ionic, for example sodium carboxymethylcellulose, hydroxyethylethylcellulose, hydroxyethylcellulose, hydroxypropylcellulose, hydroxyethylmethylcellulose and hydroxypropylmethylcellulose.
    Cellulose ethers are used in additives in such diverse applications as food, paint, oil recovery, paper, cosmetics, pharmaceuticals, adhesives, printing, agriculture, ceramics, textiles, detergents and building materials. Cellulose ethers improve the product quality in these applications and act as thickeners, water retains, suspending aids, protective colloids, film formers or thermoplastics in such various products as dispersion paints, drilling muds, ice creams, tablet coatings, wallpaper adhesives and brick adhesives.
    Nonionic cellulose ethers, such as methylcellulose, hydroxypropylmethylcellulose (also referred to as hypromellose) and methylhydroxyethylcellulose, are widely used in the pharmaceutical industry because of their ability to leach, bind and retain water, as well as to emulsify and suspend particles and form particles. Further information regarding non-ionic cellulose ethers can be found, for example, in WO92 / 09307. The peptide structure of oxytocin makes it susceptible to degradation and it is a well known fact that pharmaceutical compositions comprising oxytocin should be stored in a cool environment (about 4 ° C) to avoid significant degradation and / or degradation. or aggregation and thus lose its biological function. For example, it is recommended that Syntocinon® (Novartis), a concentrated aqueous solution of oxytocin for injection / infusion, should normally be stored at 2-8 ° C. The shelf life of 25 ° C for this product is limited to 3 months. Another example is Syntocinon® nasal spray. which should also be stored at 2-8 ° C. These aqueous formulations contain buffers, preservatives, acids, salts, co-solvents and other water-soluble additives but no gelling agents.
    Previous formulations of oxytocin comprising the ionic gelling agent sodium carboxymethyloellulose (CMC) have been found not to provide a suitable environment for oxytocin. This is illustrated, for example, in the experimental part of the present application.
    Accordingly, given the problems associated with prior pharmaceutical compositions including oxytocin, there is a need in the art to overcome or at least reduce some of the disadvantages in the art by providing pharmaceutical compositions in which the stability of oxytocin is improved. Such pharmaceutical compositions will provide for more convenient storage, such as at room temperature, and should also maintain its biological activity for an extended period of time.
    SUMMARY OF THE INVENTION The problems presented above have now been solved by the present invention, which is further explained herein. Accordingly, the present invention relates to novel pharmaceutical compositions comprising oxytocin, and / or one or more fragments and / or variant (s) thereof shown herein, as well as pharmaceutically acceptable salts thereof, and at least one non-ionic cellulose ether, as shown herein, wherein said pharmaceutical composition has a pH in the range of between about 3 and 4.
    It has surprisingly been found that a neutral, non-ionic cellulose ether, such as hydroxypropyl methyloellulose (HPMC), significantly improves the stability of oxytocin in a pharmaceutical composition, thereby providing a pharmaceutical composition which is excellent for pharmaceutical use. . This stabilizing effect of the non-ionic cellulose ether is further enhanced by the fact that said pharmaceutical composition has a low pH, such as in the range of between about 3 and 4, such as in the range of between about 3 and 3.5. This makes it possible to store oxytocin at room temperature for longer periods of time than was previously possible, when present in such a pharmaceutical composition. A favorable environment for the oxytocin peptide in such a pharmaceutical composition should also positively affect its biological activity thereby enhancing its therapeutic effect as more biologically active substance is present therein for a longer time.
    Accordingly, the present invention relates to pharmaceutical compositions comprising oxytocin (SEQ ID NO: 1), and / or one or more fragments and / or variant (s) thereof according to SEQ ID NO: 2, as well as pharmaceutically acceptable salts thereof and at least a non-ionic cellulose ether, such as hydroxypropylmethylcellulose (HPMC), wherein said pharmaceutical composition has a pH in the range of between about 3 and 4, such as about 3 and 3.5 or as otherwise exemplified herein, wherein the stability of oxytocin and / or one or more fragment your fragments and / or variants thereof have been improved. The ability of a non-ionic cellulose ether, such as HPMC, to stabilize oxytocin is not previously known and is highly desirable as there has long been a need in the art to provide pharmaceutical compositions comprising oxytocin which do not always need to be stored in a cool environment and which therefore becomes easier to handle.
    The present invention also relates to medicinal uses of a pharmaceutical composition as disclosed herein, such as in the treatment of vaginal atrophy. Accordingly, in other aspects, the present invention also relates to the use of oxytocin, and / or one or more fragments and / or variant (s) thereof, and at least one nonionic cellulose ether as shown herein, to the action of a medicament, wherein said medicament has a pH in the range of between about 3 and 4 for the treatment of a menopausal disorder, such as vaginal atro fi. In addition, the present invention relates to a method of preparing said pharmaceutical composition, as well as to methods of treating a patient with a pharmaceutical composition in accordance with the invention. 10 15 20 25 30 35 536 091 PS54264EN00 DETAILED DESCRIPTION Definitions Whenever "oxytocin", "oxytocin peptide" and / or oxytocin molecule are referred to herein, this includes oxytocin (SEQ ID NO: 1) and / or one or more fragments and / or variant (s) thereof as defined herein in accordance with the general formula SEQ ID NO: 2 or any other variant and / or fragment mentioned herein, as well as analogues and / or homologues thereof. Whenever a fragment, variant or homologue of an oxytocin molecule / peptide is referred to, it is to be understood that such variant, fragment or homologue possesses a biological activity comparable to the oxytocin molecule itself (SEQ ID NO: 1). As an example, it can be shown that a substance has oxytocin activity by performing tests that show the activity of the substance in question, for example by performing a double-blind crossed randomized protocol as described in WO0178758 (Example 1). Accordingly, a "variant" of oxytocin, referred to herein, refers to a peptide which has been varied in its amino acid structure compared to the oxytocin molecule in such a way that certain amino acid positions may have been altered by introducing other amino acids into such positions, such as natural or non-natural. natural amino acids exemplified herein or it may have been extended by adding one or more natural or non-natural amino acids to any of the ends of the peptide. In addition to this, other structural variations may also have been made to the present peptides referred to herein, such as synthetic modifications. Said "variant" still retains a biological activity similar to oxytocin and said oxytocin variant is also stabilized by being present in a pharmaceutical composition in accordance with the present invention.
    Furthermore, a "fragment" of oxytocin referred to herein is a peptide which comprises a portion of the amino acid sequence of oxytocin but where one or more amino acids may have been removed from one or both of the amino acid terminal end (s). This term also refers to a fragment of an oxytocin variant as defined in SEQ ID NO: 2, which consequently means that also encompassed by the invention is any fragment of a peptide presented by SEQ ID NO: 2.
    A "pH adjusting agent" is any agent, such as a surfactant, such as an aqueous liquid, present in a pharmaceutical composition in accordance with the present invention, which can regulate and / or maintaining the pH of said pharmaceutical composition, wherein said pH is maintained approximately within a selected range, which selected range is exemplified herein. Such a pH adjusting agent may be, for example, a buffer, such as a citrate, lactate or a phosphate buffer. A "buffer" is an ionic compound, usually a salt of a weak acid or base, which is added to a solution to resist changes in its acidity or alkalinity and thus stabilize its pH. A buffer solution is a solution containing such a compound. Other examples of pH adjusting agents are organic and inorganic acids and bases such as acetic acid, citric acid, phosphoric acid and hydrochloric acid and sodium hydroxide.
    Oxytocin is known to suffer from problems due to degradation and / or aggregation which makes it preferable to store pharmaceutical compositions comprising oxytocin in a cool environment to avoid losing its biological activity. Surprisingly, it has now been found that it is possible to prepare pharmaceutical compositions comprising oxytocin and / or one or more fragments and / or variant (s) thereof. as illustrated in SEQ ID NO: 1 and SEQ ID NO: 2, with improved stability by using at least one non-ionic cellulose ether as a pharmaceutical carrier for said oxytocin molecule, in combination with a low pH.
    When referring to an improved stability of said pharmaceutical composition, it is the stability of the biological substance, i.e. oxytocin and / or one or more fragments and / or variant (s) thereof as defined herein, which is improved. Accordingly, the biological activity and therapeutic effect of a pharmaceutical composition presented herein should be improved, by avoiding too much degradation and / or aggregation or other structural change of the oxytocin substance.
    Without being bound by any particular theory, the nonionic cellulose ether, which is an uncharged molecule, seems to stabilize the oxytocin molecule by avoiding unnecessary interference with it. Furthermore, the use of a suitable pH contributes. such as a pH in the range of between about 3 and 4, as in the range of between about 3 and 3.5, further to the stabilizing effect of the nonionic cellulose ether of oxytocin. Consequently, without again being bound by a specific theory, the low pH not only seems to stabilize oxytocin itself, but is also favorable for the non-ionic cellulose ether part of the composition. In combination, this provides an excellent composition for pharmaceutical use, wherein the biological and / or therapeutic activity of oxytocin and / or a variant and / or a fragment thereof as exemplified herein should be increased and prolonged. This pharmaceutical composition is also easier to handle for the user because due to its improved stability it does not always have to be stored in a cool environment but instead can more conveniently be kept at room temperature during use and at the same time retain its biological activity.
    The stabilizing effect of a non-ionic cellulose ether on the oxytocin molecule in a pharmaceutical composition in accordance with the present invention is in contrast to other pharmaceutical carriers used, such as sodium carboxymethylcellulose (CMC), in which formulation oxytocin has been found to degrade more rapidly and / or aggregate (see for example the comparative examples in the experimental part). Without being bound by a specific theory, this could be due to the fact that the CMC polymer is negatively charged and can be considered a polyanion, which could interact inappropriately with the oxytocin peptide.
    Accordingly, the present invention relates to a pharmaceutical composition comprising: a. Oxytocin (SEQ ID NO: 1), and / or one or more fragments and / or variant (s) thereof. In accordance with SEQ ID NO: 2, both as pharmaceutically acceptable salts thereof, and b. at least one non-ionic cellulose ether; wherein SEQ ID NO: 2 is X, -X2-X3-X4-Asn-Cys-Xs-X fl-Xy-XB-NH, wherein X1 is selected from the group consisting of Cys and none; X2 is selected from the group consisting of Tyr, Phe, and nothing; Xa is selected from the group consisting of Ile, Val, Hoph, Phe, Cha, and nothing; X 4 is selected from the group consisting of Gln, Ser, Thr, Cit, Arg, and Daba; X5 is selected from the group consisting of Pro and none; X5 is selected from the group consisting of Ile, Leu, Nothing, Val, Hos, Daba, Thr, Arg, and Cit; X 1 is selected from the group consisting of Gly, none, and Ala; X, selected from the group consisting of Gly and nothing; Wherein said pharmaceutical composition has a pH in the range of between about 3 and about 4, 1 an aspect of the invention, when X 1 is Cys a disulfide bridge is formed between X 1 and Cys.
    Accordingly, it is to be understood that when X1i of formula (I) is cysteine (Cys), the thiol group of X1 may form a disul fi d with the thiol group of said cysteine placed between asparagine (Asn) and X5 thereby forming a cyclic structure of formula ( In a further aspect, there is provided a pharmaceutical composition comprising oxytocin (SEQ ID NO: 1) and hydroxypropylmethylcellulose, and wherein the pH of said pharmaceutical is: cys-xz-xa-n-Asn-cys-xs-xo-xfxs-NH 2 composition is in the range of between about 3 and 4, such as between about 3 and 3.5 or as otherwise exemplified herein. In a further aspect, the present invention also relates to a pharmaceutical composition consisting of a non-ionic cellulose ether and oxytoxin (SEQ ID NO: 1), and / or a variant and / or a fragment thereof presented herein (SEQ ID NO: 1). 2), as well as pharmaceutically acceptable salts thereof, and a pH adjusting agent, wherein said pharmaceutical composition has a pH in the range of between about 3 and 4, such as in the range of about 3 and about 3.5. In one aspect, said pharmaceutical composition consists of oxytocin (SEQ ID NO: 1), hydroxypropyl methylcellulose (HPMC), and a pH adjusting agent, wherein said pharmaceutical composition has a pH in the range of between about 3 and 4, such as between about 3 and about 3.5 or as otherwise exemplified herein. In one aspect, said pH adjusting agent is a buffer, such as a citrate or a lactated buffer.
    In one aspect of the present invention, said pharmaceutical composition comprises oxytocin, i.e. when X 1 is Cys, X 2 is Tyr, X 3 is Ile, X 1 is Gln, X 5 is Pro, X 1, is Leu, X 7 is Gly, and X8 is not in SEQ ID NO: 2, (Cys-Tyr-Ile-Gln-Asn-Cys-Pro-Leu-Gly-NHz) (SEQ ID NO: 1). The variant (s) and / or fragments of oxytocin in said pharmaceutical composition (SEQ ID NO: 2) may also be selected from the group consisting of the following compounds, as well as pharmaceutically acceptable salts thereof. : Mesotocin: Cys-Tyr-IIe-GIn-Asn-Cys-Pro-Ile-Gly-NH2 (SEQ ID NO: 3) X1 is Cys, X2 is Tyr, X5 is Ile, X4 is Gln, X5 is Pro, X5 is Ile, X1 is Gly, and X5 is none; Isotocin: Cys-Tyr-IIe-Ser-Asn-Cys-Pro-Ile-Gly-NH2 (SEQ ID NO: 4) X1 is Cys, X2 is Tyr, X5 is Ile, X4 is Ser, X5 is Pro, X5 is Ile, X1 is Gly, and X1, is nothing; Annetocin: Cys-Phe-VaI-Arg-Asn-Cys-Pro-Thr-Gly-NH 2 (SEQ ID NO: 5) X 1 is Cys, X 2 is Phe, X 5 is Val, X 4 is Arg. X5 is Pro, X5 is Thr, X1 is Gly, and X5 is none; Vasotocin: Cys-Tyr-Ile-Gln-Asn-Cys-Pro-Arg-Gly-NH2 (SEQ ID NO: 6) X1 is Cys, X2 is Tyr, X5 is Ile, X4 is Gln, X5 is Pro, X5 is Angry, X1 is Gly, and X5 is none; Vasopressin: Cys-Tyr-Phe-Gln-Asn-Cys-ProArg-Gly-NH2 (SEQ ID NO: 7) X1 is Cys, X2 is Tyr, X5 is Phe, X4 is Gln, X5 is Pro, X5 is Arg, X1 is Gly, and X5 is none; Cys-Tyr-Ile-Gln-Asn-Cys-NH2 (SEQ ID NO: 8) X1 is Cys, X2 is Tyr, X5 is Ile, X4 is Gln, and X5-X5 is none; Cys-Tyr-Ile-Gln-Asn-Cys-Pro-NH2 (SEQ ID NO: 9) X1 is Cys, X2 is Tyr, X5 is Ile, X4 is Gln, X5 is Pro, and Xe-X; year nothing; Cys-Tyr-III-Gln-Asn-Cys-Pro-Leu-NH2 (SEQ ID NO: 10) X1 is Cys, X2 is Tyr, X5 is Ile, X4 is Gln, X5 is Pro, X5 is Leu, and X1 .5 is nothing; Tyr-Ile-Gln-Asn-Cys-Pro-Leu-Gly-NH2 (SEQ ID NO: 11) X1 is none, X2 is Tyr, X5 is Ile, X4 is Gln, X5 is Pro, X5 is Leu, X1 is Gly, and X5 is nothing; IIe-Gln-Asn-Cys-Pro-Leu-Gly-NH2 (SEQ ID NO: 12) X1-X2 is none, X5 is Ile, X4 is Gln, X5 is Pro, X5 is Leu, X1 is Gly, and X5 is nothing; Gln-Asn-Cys-Pro-Leu-Gly-NH2 (SEQ ID NO: 13) 10 15 20 25 30 35 536 091 PS54264SE00 10 X1-X5 is none, X4 is Gln, X5 is Pro, X5 is Leu, X1 is GIy, and X5 is nothing; IIe-GIn-Asn-Cys-Pro-NH5 (SEQ ID NO: 14) X1-X5 is none, X5 is Ile, X4 is Gln, X5 is Pro, and X5-X5 is none; Cys-Tyr-Ile-Gln-Asn-Cys-Pro-Leu-GIy-GIy-NH; (SEQ ID NO: 15) X1 is Cys, X5 is Tyr, X5 is Ile, X4 is Gln, X5 is Pro, X5 is Leu, X1 is Gly, and X5 is Gly; Gln-Asn-Cys-Pro-Leu-Leu-NHZ (SEQ ID NO: 16) X1-X5 is none, X4 is Gln, X5 is Pro, X5 is Leu, X1 is Leu, and X5 is none; Cys-Tyr-VaI-Thr-Asn-Cys-Pro-Leu-GIy-NH5 (SEQ ID NO: 17) X1 is Cys, X5 is Tyr, X5 is Val, X4 is Thr, X5 is Pro, X5 is Leu, X1 is Gly, and X5 is none; Cys-Tyr-Hoph-Thr-Asn-Cys-Pro-VaI-Gly-NH, (SEQ ID NO: 18) X1 is Cys, X2 is Tyr, X5 is Hoph, X4 is Thr, X5 is Pro, X5 is Val , X1 is Gly, X5 is none; Cys-Tyr-Phe-Cit-Asn-Cys-Pro-Leu-GIy-NH2 (SEQ ID NO: 19) X1 is Cys, X5 is Tyr, X5 is Phe, X4 is Cit, X5 is Pro, X5 is Leu, X1 is Gly, and X5 is none; Cys-Tyr-Cha-Arg-Asn-Cys-Pro-Hos-Ala-NHZ (SEQ ID NO: 20) X1 is Cys, X5 is Tyr, X5 is Cha, X4 is Arg, X5 is Pro, X5 is Hos, X1 is Ala, and X5 is none; Cys-Tyr-VaI-Daba-Asn-Cys-Pro-Daba-AIa-NH, (SEQ ID NO: 21) X1 is Cys, X5 is Tyr, X5 is Val, X4 is Daba, X5 is Pro, X5 is Cit , X1 is Ala, and X5 is none; Cys-Tyr-Hoph-Daba-Asn-Cys-Pro-Cit-AIa-NH5 (SEQ ID NO: 22) X1 is Cys, X5 is Tyr, X5 is Hoph, X4 is Daba, X5 is Pro, X5 is Cit, X1 is Ala, and X5 is none; Cys-Tyr-Phe-Arg-Asn-Cys-Pro-VaI-Ala-NHZ (SEQ ID NO: 23) X1 is Cys, X5 is Tyr, X5 is Phe, X4 is Arg, X5 is Pro, X5 is Val, X1 is Ala, and X5 is none; and Cys-Tyr-Cha-Cit-Asn-Cys-Pro-Arg-Gly-NH, (SEQ ID NOs24) 536 9 '| PS54264SE00 11 X1 is Cys, X2 is Tyr, X3 is Cha, X4 is Cit, X5 is Pro, X5 is Arg, X7 is Gly, X; is nothing; Accordingly, in one aspect, the present invention relates to a pharmaceutical composition, wherein said one or more fragments and / or varlant (s) of oxytocin are selected from the group consisting of the peptides corresponding to SEQ ID NOzâ-SEQ ID NO: 24.
    The non-natural amino acids of said substances have the following structures: Cyclohexylalanine, referred to herein as Cha, O NH, Homophenylalanine, referred to herein as Hoph, on NH, Citrulline, herein referred to as Cit, O i., .. _. / <0 H OH NH, 1 5 Diaminobutanoic acid, herein referred to as Daba, and 536 G91 PS54264SEO0 12 WN oH nu, Homoserine, herein referred to as Hos, O H0 NH When a position in SEQ ID NO: 2 is referred to as "nothing", it means that it represents a single bond between the items (letter, atom or group).
    Other variants of oxytocin may also be used in the pharmaceutical compositions according to the present invention, such as naturally occurring or artificially modified variants, analogs and / or derivatives of oxytocin, mesotocin, isotocin and / or annetocin. Such variants can be obtained by the addition, insertion, elimination or substitution of at least one amino acid in these hormones. Other substances include precursors, metabolites, such as metabolic derivatives, for example metabolic degradation products, agonists or analogs of the substances mentioned herein which exhibit the same properties. Metabolic derivative or metabolic degradation products may be oxytocin-like peptides, for example with nine amino acids such as oxytocin, mesoticin, isotocin and annetocin from which one or more amino acids have been removed from either the carboxy-terminal or amino-terminal terminal or both carboxy-terminal terminal and both carboxy-terminal terminal from each terminal. In some respects, one, two or three amino acids may have been removed from the carboxy terminus, for example Gly alone, Gly and Leu or Gly, Leu and Pro. Preferably, one, two or three amino acids may have been removed from the amino terminus, for example only Cys, Cys and Tyr or Cys, Tyr and lle. In certain aspects, one, two or three amino acids may have been removed both from the carboxy terminal end, i.e., only Gly, Gly and Leu or Gly, Leu, and Pro, and one, two or three amino acids may have been removed from the amino terminus, i.e. only Cys, Cys and Tyr or Cys, Tyr and lle. It can be determined that these variants are analogues of oxytocin, mesotocin, isotocin or annetocin by immunological methods, for example RIA (radioimmunoassay), IRMA (radiometric methods). RIST (radioimmunosorbent test) and RAST (radloallergosorbent test). The invention also includes variants of oxytocin having at least 50, 60, 70, 80, 90, 95, 96, 97, 98 or 99% sequence identity with oxytocin, said variants exhibiting an oxytocin activity as defined herein.
    Annetocin has been isolated from the earthworm, as described in Oumi T. Ukena K, Matsushima 0, lkeda T, Fujita T, Minakata H, Nomoto K, Annetocin: an oxytocin-related peptide isolated from the earthworm, Eisenia foetida, Biochem Biophys Res Commun 1994 , January 14; 198 (1); 393-399.
    There is also an opportunity to create new compounds with oxytocin activity through computer simulation. Methods for computer simulation are known to those skilled in the art, for example, as described in EP0660210 A2.
    The invention also relates to a pharmaceutical composition comprising oxytocin or a variant thereof in both D- and L-forms, as well as the racemate thereof. In some aspects, the upturn refers to the L-shape. By inversion of the peptide sequence thereof, the D-form can be converted to the L-form. These and the above peptides can be produced by methods known to those skilled in the art, for example according to Merrifield, P. B., "Solid Phase Synthesis", Angew. Chemistry, 1985, no. 97, pp. 801.
    The phenolic compositions according to the invention may in some cases contain substances which increase or enhance the effects of oxytocin. Such substances could increase the release of oxytocin and / or the moiety or affinity of oxytocin receptors, such as estrogen or drugs that have an alpha-agonist effect, such as clonidine. It should be noted that pharmaceutically acceptable salts of the compounds according to the invention are included within the scope of the invention. Examples of salts of the compounds are pharmaceutically acceptable acid and base addition salts.
    The term "pharmaceutically acceptable acid addition salts" is intended to be any non-toxic organic or inorganic acid addition salt of the compounds of SEQ ID NO: 2, any other variant (s) and / or fragment of oxytocin as described herein.
    Examples of illustrative inorganic acids which form suitable salts are hydrochloric acid, hydrobromic acid, sulfuric acid, phosphoric acid and acidic metal salts such as sodium monohydrogen orthophosphate and potassium hydrogen sulphate. Examples of illustrative organic acids which form suitable salts are mono-, di- and tricarboxylic acids. Examples of such acids are acetic acid, glycolic acid, lactic acid, pyruvic acid, malonic acid, succinic acid. glutaric acid, fumaric acid, malic acid, tartaric acid, citric acid, ascorbic acid, maleic acid, hydroxymaleic acid, benzoic acid, hydroxybenzoic acid, phenylacetic acid, cinnamic acid, salicylic acid, 2-phenoxybenzoic acid, and sulfonic acids such as p-toluenesulfonic acid, and 2-hydroxysulfonic acid. Such salts may be either in hydrogenated or in anhydrated form.
    The acidic addition salts of these compounds are generally water-soluble and various hydro-organic solvents, and which, in comparison with the free base forms thereof, generally exhibit higher melting points.
    The term "pharmaceutically acceptable basic addition salts" is intended to be any non-toxic organic or inorganic basic addition salts of the compounds of SEQ ID NO: 2, and / or any other variant (s) and / or fragment of oxytocin described herein. Examples of illustrative inorganic bases which form suitable salts are alkaline and alkaline earth metal hydroxides and carbonates such as sodium hydroxide, sodium carbonate, potassium hydroxide, potassium carbonate, calcium hydroxide, calcium carbonate, magnesium hydroxide, magnesium carbonate and ammonia. and picoline.Either mono- or dibasic salts can be formed with such compounds.The basic addition salts of these compounds are generally water soluble and various hydro-organic solvents, and which, compared with the free base forms thereof, generally exhibit higher melting points.
    The nonionic cellulose ethers of the pharmaceutical compositions of the present invention are based on cellulose, which has been chemically modified to achieve solubility in water by substituting various groups on the cellulose backbone. The substituents are characterized in that they have no electric charge when dissolved in water at a neutral pH.
    The properties of the nonionic cellulose ethers are determined by molecular weight, (for example degree of polymerization), the type of substituents and also by the number and distribution of the substituents along the molecule. Accordingly, depending on the oxytocin and / or fragment and / or variant thereof which is present in the pharmaceutical composition, the nonionic cellulose ethers may have different properties and will be determined depending on the prevailing circumstances.
    The cellulose ethers in accordance with the present invention are non-ionic, wherein alkyl and / or hydroxyalkyl groups are attached to the anhydroglucose moieties through ether linkages, which form hydroxyalkylalkylcelluloses, wherein the alkyl groups have from 1 to 4 carbon atoms.
    Representative cellulose ethers for use in the pharmaceutical compositions according to the present invention are methylcellulose (MC), hydroxyethylmethylcellulose (HEMC), hydroxypropylmethylcellulose (HPMC), hydroxyethylethylcellulose (HEEC), and hydroxypropylcellulose (HPC). These polymers have substituents which are either non-polar (for example methyl) or slightly polar (for example hydroxyethyl), which in combination with the hydrophilic cellulose backbone provide an amyl polymer.
    Accordingly, the present invention relates to pharmaceutical compositions wherein the at least one non-ionic cellulose ether is selected from one or more of the following cellulose ethers: MC, HPMC, HEEC, HEMC and / or HPC or as further exemplified herein. In one aspect of a pharmaceutical composition according to the present invention, said nonionic cellulose ether is hydroxypropyl methylcellulose (HPMC). The pH of a pharmaceutical composition in accordance with the present invention, and applicable to all embodiments and aspects thereof as defined herein may be in the range between about 3 and 4.5, such as between about 3.5 and about 4.5, about 3.5 and 4, about 3 and 4, about 3 and 3.5, about 3 and 3.1, about 3 and 3.2, about 3 and 3.3, about 3 and 3.4, about 3 and 3.6, about 3 and 3.7, about 3 and 3.8, about 3 and 3.9, about 3.1 and 3.2, about 3.1 and 3.3, about 3.1 and 3.4, about 3 , 1 and 3.5, about 3.1 and 3.6, about 3.1 and 3.7, about 3.1 and 3.8, about 3.1 and 3.9, 10 15 20 25 30 536 091 PS54264SE00 16 about 3.2 and 3.3, about 3.2 and 3.4, about 3.2 and 3.5, about 3.2 and 3.6, about 3.2 and 3.7, about 3.2 and 3.8, about 3.2 and 3.9, about 3.3 and 3.5, about 3.3 and 3.6, about 3.3 and 3.7, about 3.3 and 3.8 or about 3, 3 and 4.
    It should be noted that these values are not exact, which means that they may vary slightly around the values given. The pH of a pharmaceutical composition as initiated herein can be controlled by adding a pH adjusting agent to said pharmaceutical composition, such as a buffer. In the context of the present invention, said buffer may be a lactate buffer, a citrate buffer, a phosphate buffer or a mixture thereof, but is not limited thereto.
    The concentration of a buffer to be added to the pharmaceutical composition according to the invention may be between about 20 to 100 mM, such as between 25 mM-100 mM or about 25-50 mM, about 25 mM to 75 nM or about 50 to 70 mM in an aqueous solution but is not limited thereto. It should be noted that these values are not exact, which means that they may vary slightly around the values given. Depending on the pH required in the pharmaceutical composition in accordance with the present invention, the concentration of the buffer may vary in accordance with the above. In one aspect of the present invention, a pharmaceutical composition comprises oxytocin (SEQ ID NO: 1) and hydroxypropyl methylcellulose (HPMC), optionally in combination with buffers, other pH adjusting agents or other components exemplified herein, such as a preservative, such as benzoic acid. In one aspect, the pH of said composition of oxytocin and HPMC is in the range of between about 3-4, such as about 3-3.5, about 3.1 to 3.6, or about 3 to 3.8, or as further defined herein. .
    In a phenotoxic composition in accordance with the present invention, the concentration of oxytocin and / or a fragment and / or a variant thereof as incorporated herein may be between about 0.1 to 1.5 mg / g, such as about 0.5 to about 1 mg. , 5 mg / g, about 0.5 to about 1 mg / g, about 0.5 to about 1.2 mg / g, about 0.2 to about 0.5 mg / g, about 0.1 to about 0 .8 mg / g or about 0.2 to about 1.2 mg / g of the total pharmaceutical composition as defined herein. In one aspect, a pharmaceutical composition according to the present invention comprises about 1 mg of oxytocin, about 2% by weight of hydroxypropyl methylcellulose in about 25 mM of citrate or lactate buffer, and optionally about 1 mg of benzoic acid. . In another aspect, a pharmaceutical composition in accordance with the present invention comprises about 0.9 mg / g oxytocin, about 1.1 mg / g benzoic acid, and about 2% by weight of hydroxypropyl methylcellulose (HPMC) in about 25 mM lactate buffer. The pH of the pharmaceutical compositions described above is in the range of between about 3 and 4, such as about 3 to about 3.5 or about 3.1 to about 3.6 or about 3.2 to 3.5 or as further exemplified herein.
    The concentration of the at least one nonionic cellulose ether, such as hydroxypropyl methylcellulose (HPMC) is provided relative to the pH adjusting agent of the pharmaceutical composition, such as a buffer presented herein, for example, provides 2% by weight of non-ionic cellulose ether in 25 mM buffer a certain viscosity due to the amount of the at least one non-ionic cellulose ether present in said buffer. Accordingly, the concentration of the non-ionic cellulose ether is presented in% by weight (vt%) in said buffer or other pH adjusting agent. In the overall pharmaceutical composition, the amount of non-ionic cellulose ether is slightly less due to the addition of the therapeutic substance (oxytocin and / or one or more fragments and / or variant (s) thereof, as shown herein) to said composition.
    The concentration of oxytocin and / or one or more fragments and / or variant (s) thereof as incorporated herein is provided relative to the total pharmaceutical composition, i.e. the buffer and the non-ionic cellulose ether, and optionally other additives such as a preservative. Accordingly, 1 mg of oxytocin means that 1 mg of oxytocin is present in 1 g of total pharmaceutical composition.
    The present invention also relates to the pharmaceutical compositions shown herein for use as medicaments. In one aspect of the invention, the invention relates to a pharmaceutical composition comprising oxytocin (SEQ ID NO: 1) and hydroxypropyl methylcellulose, optionally in combination with buffers, other pH adjusting agents and / or additional components exemplified herein, such as a preservative such as benzoic acid , for use as a medicine. Such a pharmaceutical composition has a pH within the ranges exemplified herein. An example of a medical use of the compositions of the present invention is for vaginal use, such as vaginal atro fi. Such a medical use of oxytocin is shown by the present inventor in WO0178758.
    For examples of other medicinal uses of oxytocin with which the pharmaceutical compositions of the present invention are useful, we refer to WO02102832, WO02067974, WO03017922, WO0018424, WO9843661, WO9843662, and WO9843660. Accordingly, the present invention also relates to a pharmaceutical composition presented herein for a medical use exemplified in any of the above-mentioned patent documents. Such medicinal uses are, for example, to create eustasis, cancer in situ and cervicitis, inmation, cell regeneration, wound healing, preference and acceptance, and for the treatment of pain, when a pharmaceutical composition according to the present invention is suitable for use in the treatment of a such a disease state, such as when a topical treatment is suitable for the treatment thereof.
    The term "eustasis" refers to a psychophysiological condition, ie a combination of a psychological and physiological condition. The psychological state is characterized by calm and positive social interactions such as trust and breastfeeding. The physiological condition is characterized by muscle relaxation, decreased cardiovascular activity and improved gastrointestinal activity. In addition to this, pulse rate and blood pressure are maintained at a low, healthy and balanced level, and the vaginally controlled gastrointestinal area is activated, favoring digestion and storage of nutrients.
    The term "in situ and cervical cancer" refers to the consequences of diseases of the vagina and cervix that originate in infections, as well as in breastfeeding. In the context of recovery, in situ cancer is related to the cervix. Such diseases include, in addition to in situ and cervical cancer, also pre-cancerous disease states, squamous cell carcinoma and coilocytosis. In situ cancer refers to a neoplastic entity in which the tumor cells are confined to the epithelium where they originate, without invasion of the basement membrane. Cervicitis refers to inflammation of the cervix uteri, ie the lower and narrower end of the uterus, between the isthmus and the ostium uteri. The epithelium of the cervix uteri is quite different from the epithelium of the rest of the uterus. Koilocytosis is a consequence of the herpes virus.
    The pharmaceutical compositions as defined herein are all intended for topical use, such as for vaginal use. When they are for vaginal use, this may be for the treatment of a menopausal condition, such as vaginal atrophy or any other condition mentioned in WO0178758. Accordingly, in one aspect, the present invention relates to a pharmaceutical composition comprising oxytocin (SEQ ID NO: 1) and hydroxypropylmethylcellulose (HPMC), optionally in combination with buters, other pH adjusting agents or additional components exemplified herein, such as benzoic acid for vaginal use. such as for the treatment of a menopausal disease, for example vaginal atrophy. p1-1 in such a pharmaceutical composition may be in the range of between about 3 to about 4, such as about 3 to about 3.5, about 3.1 to about 3.6 or about 3.1 to about 3.8, or as otherwise exemplified herein. In all aspects of the present invention, it is to be understood that whenever reference is made to a particular medicinal use of said pharmaceutical composition, such as in the use for treating vaginal atro fi, this also refers to the use of oxytocin and / or one or more fragments and / or variants thereof, such as those introduced herein, and at least one non-ionic cellulose ether in the manufacture of a medicament for the treatment of certain diseases, as exemplified herein, wherein said medicament has a pH in the range of between about 3 and 4, such as about 3 and 3 , 5 or as further exemplified herein.
    As shown herein, the presence of at least one non-ionic cellulose ether in the pharmaceutical composition effectively improves the stability of the oxytocin molecule therein.
    Accordingly, the present invention also relates to the use of at least one non-ionic cellulose ether to stabilize, alternatively improve the stability of a pharmaceutical compound comprising oxytocin (SEQ ID NO: 1) and / or one or more fragments and / or variant (s). of oxytocin, as defined in SEQ ID NO: 2 or otherwise exemplified herein. In one aspect, the invention relates to the use of the at least one nonionic cellulose ether hydroxypropylmethylcellulose (HPMC) to stabilize, alternatively improve the stability of a pharmaceutical composition comprising oxytocin (SEQ ID NO: 1) and / or a fragment and / or a variant of oxytocin, as well as pharmaceutically acceptable salts thereof as defined in SEQ ID NO: 2 or as otherwise defined herein, wherein said pharmaceutical composition optionally also comprises a buffer, other pH-regulating agents or additional components exemplified herein, such as a preservative. As previously mentioned herein, the combination of using a low pH in the pharmaceutical composition further enhances the stabilizing effect of the at least one nonionic cellulose ether. Without being bound by a specific theory, the at least one non-ionic cellulose ether appears to stabilize the oxytocin molecule and / or one or more fragments and / or variant (s) as defined herein by making it less prone to degrade and / or aggregate or otherwise structurally altered, possibly by not interfering with the oxytocin molecule.
    Whenever reference is made to an improved stability / stabilizing effect herein, this may mean the reduction of aggregation and / or degradation or other structural changes of the oxytocin molecule in the pharmaceutical composition defined herein, but is bound by this particular theory. According to such a theory, a reduction in aggregation and / or degradation or other structural changes is associated with an improved stability of the oxytocin molecule. The improvement in stability of a pharmaceutical composition comprising at least one non-ionic cellulose ether is further illustrated in the experimental part including comparative examples with the CMC gel. In another aspect, the present invention relates to a method of preparing a pharmaceutical composition with improved stability comprising the steps of: a) b) preparing a buffer solution and adjusting the pH therein, adding oxytocin and / or one or more fragments and / or or variant (s) thereof according to SEQ ID NO: 2 or as otherwise specified herein, as well as pharmaceutically acceptable salts thereof to said buffer, and adding at least one non-ionic cellulose ether to the solution obtained in step b); wherein optionally a preservative, such as benzoic acid, is added to the buffer solution in step a) before oxytocin and / or one or more fragments and / or variant (s) thereof according to SEQ ID NO: 2 or as otherwise exemplified herein is added to said buffer solution and wherein the pH of said pharmaceutical composition is adjusted to fall in the range of between about 3 and about 4, such as in the range of about 3 and about 3.5 or as otherwise exemplified herein. After step c), the solution is allowed to rest for a while to obtain a viscous solution or a gel. Typically, said viscous solution or gel is obtained after about 1-2 0) days. Said method may also in certain aspects comprise the use of pH adjusting agents other than buffers, such as organic and inorganic acids and bases, such as acetic acid, citric acid, phosphoric acid and hydrochloric acid and sodium hydroxide, which accordingly used in step a) of said method.
    In the present method, what is meant is that the stability of oxytocin, and / or one or more of the fragments and / or variant (s) thereof mentioned herein is enhanced by preparing the composition together with the at least one non-ionic cellulose ether. In one aspect of the present method, it is at least one nonionic cellulose ether hydroxypropyl methylcellulose (HPMC). In one aspect, the method relates to the preparation of a pharmaceutical composition comprising oxytocin (SEQ ID NO: 1) and hydroxypropylmethyloellulose (HPMC).
    Administration of an oxytocin molecule may in some respects benefit from adding touch, pressure, massage, heat or infrared light to the skin, leading to improved skin permeability. Hirvonen, J., Kalla, Y N, and Gay, R H. Transdermal delivery of peptides by iontophoresis. Nat Biotechnol December 1996; 14 (13): 1710-1713 describes how the transport of a drug can be improved via the skin by using the driving force of an applied electric field. lontophoresis can be achieved at a slightly alkaline pH.
    There are various processes described for the synthetic production of oxytocin; commercial processes are described, for example, in U.S. Pat. 2,938,991 and 3,076,797. In one aspect, the present invention relates to a pharmaceutical composition comprising a. oxytocin (SEQ ID NO: 1), and / or one or more fragments and / or variant (s) thereof according to SEQ ID NO: 2, as well as pharmaceutically acceptable salts thereof, and b. at least one non-ionic cellulose ether; wherein SEQ ID NO: 2 is X1-X2-X3-X4-Asn-Cys-X5-Xe-Xy-Xg-NH2 wherein X1 is selected from the group consisting of Cys and none; X2 is selected from the group consisting of Tyr, Phe, and nothing; Xa is selected from the group consisting of lle, Val, Hoph, Phe, Cha, and nothing; X., is selected from the group consisting of Gln, Ser, Thr, Cit, Arg, and Daba; X5 is selected from the group consisting of Pro and none; 10 15 20 25 30 536 091 PS54264EN00 22 X6 is selected from the group consisting of lle, Leu, nothing, Val, Hos, Daba, Thr, Arg, and Cit; X7 is selected from the group consisting of Gly, Nothing, and Ala; X8 selected from the group consisting of Gly and none; wherein oxytocin, and / or said one or more fragments and / or variant (s) thereof are present in an amount of about 0.1 to 1.5 mg / g, such as 0.5 to about 1.5 mg / g, about 0.5 to about img / g, about 0.5 to about 1.2 mg / g, about 0.2 to about 0.5 mg / g, about 0.1 to about 0.8 mg / g, or about 0.2 to about 1.2 mg / g of the total composition, and wherein said at least one non-ionic cellulose ether is present in an amount of about 1-3% by weight, such as 2% by weight in a pH adjusting agent, such as a buffer, said buffer having a concentration of about 25 mM to about 100 mM. In one aspect, the at least one nonionic cellulose ether is hydroxypropylmethylcellulose. In some aspects, the pharmaceutical compositions of the present invention may optionally contain other components, such as inert components or components having a physical or biological function. In some aspects, an antibiotic or analgesic is also present in a pharmaceutical composition in accordance with the invention.
    The invention also relates to a method of treating a menopausal disorder, such as vaginal atrophy, in a patient in need thereof comprising administering an appropriate amount of a pharmaceutical composition comprising a. Oxytocin (SEQ ID NO: 1), and / or one or more fragment your fragments and / or variant (s) thereof according to SEQ ID NO: 2, as well as pharmaceutically acceptable salts thereof, and b. at least one non-ionic cellulose ether; to a patient in need thereof, wherein the pH of said pharmaceutical composition is in the range of between about 3 and 4 or as otherwise exemplified herein. Said pharmaceutical composition for use in a method of treating a menopausal disorder may be any pharmaceutical composition shown herein. In one aspect, said pharmaceutical composition administered to a patient in need thereof comprises oxytocin and hydroxypropyl methylcellulose (HPMC). The present invention also relates to a method of treating any disease or disorder previously mentioned herein comprising administering an appropriate amount of a pharmaceutical composition comprising a. Oxytocin (SEQ ID NO: 6). : 1), and / or one or more fragments and / or variant (s) thereof according to SEQ ID NO: 2, as well as pharmaceutically acceptable salts thereof, and b. At least one non-ionic cellulose ether; to a patient in need thereof, wherein the pH of said pharmaceutical composition is in the range of between about 3 and 4 or as otherwise exemplified herein. Said pharmaceutical composition for use in a method of treating a disease or disorder previously mentioned herein may be any pharmaceutical composition shown herein. In one aspect, said pharmaceutical composition administered to a patient in need thereof comprises oxytocin and hydroxypropylmethylcellulose (HPMC). In another aspect, the present invention relates to a pharmaceutical composition comprising about 1 mg / g of oxytocin, about 2% by weight of hydroxypropyl methylcellulose (HPMC) in about 25 mM citrate or lactate buffer and optionally about 1 mg / g of benzoic acid, and wherein the pH in said composition is about 3. In another aspect, the present invention relates to a pharmaceutical composition comprising about 1 mg / g oxytocin, about 2% by weight of hydroxypropyl methylcellulose (HPMC) in about 25 mM lactate buffer and wherein the pH of said composition is about 3.8. In another aspect, the present invention relates to a pharmaceutical composition comprising about 0.9 mg / g oxytocin, about 1.1 mg benzoic acid, 2% by weight of hydroxypropyl methylcellulose (HPMC) in about 25 mM lactate buffer and wherein the pH of said composition is about 3.8. In another aspect, the present invention relates to a pharmaceutical composition comprising oxytocin, hydroxypropyl methylcellulose, a citrate or lactate buffer, and optionally benzoic acid. In one aspect, said pharmaceutical composition comprises about: 1 mg / g oxytocin, about 2% by weight of hydroxypropyl methylcellulose in about 25 mM citrate or lactate buffer, and optionally about 1 mg / g benzoic acid. In another aspect, the present invention relates to a pharmaceutical composition comprising about: 1 mg / g oxytocin, 2% by weight of hydroxypropyl methylcellulose in a 25 mM lactate buffer, and optionally 1 mg / g benzoic acid.
    The present invention also relates to a kit comprising a pharmaceutical composition comprising: a. Oxytocin (SEQ ID NO: 1), and / or one or more fragments and / or variant (s) thereof according to SEQ ID NO: 2, both as pharmaceutically acceptable salts thereof, and b. at least one non-ionic cellulose ether; and a container for said pharmaceutical composition optionally in combination with instructions for use of said pharmaceutical composition, wherein the pH of said pharmaceutical composition is in the range of between about 3 and 4 or as otherwise exemplified herein. In a kit presented herein, any pharmaceutical composition described herein may be used. Said container present in said kit may, for example, be in the form of a plastic tube.
    The following invention is illustrated in the following experimental section but is not intended to be limited thereto.
    Experimental part Stressed stability studies with oxytocin in citrate / lactate buffers with Na-CMC / HPMC at 70 ° C Summary The stability of oxytocin in citrate buffer with or without benzoic acid and with or without Na-CMC was studied at 70 ° C for two weeks.
    In another set of samples, the stability of oxytocin in citrate buffer with or without benzoic acid and with or without HPMC (Hypromellose or hydroxypropylmethylcellulose) was studied at 70 ° C for three weeks.
    In a third set of samples, the stability of oxytocin in lactate buffer with HPMC and with or without benzoic acid at 70 ° C for two weeks was studied. Background The stability of oxytocin in aqueous solutions of different pH has been studied previously. In this study, it was concluded that the best stability was obtained at pH 4.5. In a later stability study, it was found that the stability of oxytocin is strongly dependent on formulation and storage conditions.
    In a third study, the stability of oxytocin in lactate buffer and citrate buffer with benzoic acid as a preservative was studied instead of the parabens used in previous studies. Parabens can cause degradation of gelling agents Na-CMCÉ However, reduced viscosity with benzoic acid was also observed.
    In the present study, the effect of the gelling agent (Na-CMC / HPMC) in citrate lactate buffer with or without benzoic acid as a preservative is studied.
    Equipment HPLC system in accordance with Ph.Eur. 2.2.29 HPLC system: Alliance 2695 from Waters, UV detection at 220 nm Column: YMC.-PAC ODS-A C18 250 * 4.6 mm, 5 pm Mobile phase: A: 15.6 g / l Sodium dihydrogen phosphate B : Acetonitrile: Water (50:50) Chromatography Software: MassLynx Version 4.1 from Water's pH Meter: Jenway 370 Chemicals Acetonitrile HPLC Grade S (ACN), Rathburn Chemicals Sterilized Water, Baxter Sodium Dihydrogen Phosphate, Merck, 6346 Oxy, 6346 Oxy Latvia Sample preparation The buffer solution was prepared by dissolving an appropriate amount of either citric acid or lactic acid in distilled water, about 90% of the total volume. The pH was adjusted to the desired value by dropwise addition of 5 M NaOH (aqueous) with magnetic stirring. Finally, the solution was transferred to a volumetric ash and the volume was adjusted to the final volume by distilled water.
    The preservative benzoic acid was weighed into a glass ash and the desired volume of buffer solution was added. After sealing the ash, benzoic acid was dissolved by treating the ash in a hot (about 50 ° C) ultrasonic bath for about 30 minutes.
    After cooling to room temperature, oxytocin was added to the clear solution and dissolved by gentle mixing. Finally, the required amount of non-ionic cellulose ether was added and the mixture was shaken by hand and then left to swell and dissolve at room temperature to give a viscous solution / gel.
    The final samples were stored in glass bottles with butyl rubber stoppers and Al cap seal.
    Sample Composition ACA101108-1 1.0 mg / l oxytocin in 100 mM citrate buffer, pH 3.1 ACA101108-2 1.2 mg / l oxytocin + 1.0 mg / l benzoic acid in 100 mM citrate buffer, pH 3.1 ACA101108-3 1.0 mg / l oxytocin + 1.0 mg / g benzoic acid + 2.6% Na-CMC in 100 mM citrate buffer, pH 3.1 ACA101108-4 1.0 mg / g oxytocin + 2.6% Na-CMC in 100 mM citrate buffer, pH 3.1 ACA101123- 1.0 mg / g oxytocin in 25 mM citrate buffer, pH 3.10 ACA101123-2 1.0 mg / g oxytocin + 1.0 mg / g benzoic acid in 25 mM citrate buffer, pH 3.10 ACA101123-3 1.0 mg / g oxytocin + 1 .0 mg / kg benzoic acid + 2.0% HPMC in 25 mM citrate buffer, pH 3.10 ACA101123-4 1.0 mg / h oxytocin + 2.0% HPMC in 25 mM citrate buffer, pH 3.10 ACA101207-1 1 mg / l oxytocin + 2 .0% HPMC in 25 mM lactate buffer, pH 3.79 ACA101207-2 0.9 mg / g oxytocin + 1.1 mg / g benzoic acid + 2.0% HPMC in 25 mM lactate buffer, pH 3.79 10 15 20 25 536 090 PS54264SE00 27 Experimental procedure The samples were stored at different temperatures and samples were removed for analysis according to the stability scheme below. Prior to analysis, 30 mg of each of the liquid samples was diluted with 1.00 mobile phase A and then injected into HPLC systems.
    Analysis All samples were visually inspected and their pH values were measured before analysis.
    HPLC Mobile Phase A: 15.6 g / l Sodium dihydrogen phosphate B: Acetonitrile water (50:50) Gradient profile: Time (min) A (%) B (%) 0 95.0 5.0 1 95.0 5.0 30.0 5 .0 95.0 40.0 5.0 95.0 45.0 95.0 5.0 Flow: 1.00 ml Injection volume: 20 μl Injection washing solution: Water: Aoetonitrile Evaluation All samples were first visually inspected and analyzed for color changes or tendencies for precipitation and changes in viscosity. The pH was then measured and the samples were further analyzed for oxytocin and benzoic acid content. Concentrations were determined against extreme standard curves for oxytocin and benzoic acid, respectively. All samples were prepared in duplicate and then analyzed by a single HPLC injection (EXP-11-AJ1634). 536 091 Pss42e4sEoo 28 Results Table 1a) -c): pH, oxylocine and benzoic acid content of the samples in series ACA101108 after up to 2 weeks of storage. ND = not determined.
    Time 0 Sample Benzoic acid Oxytocln pH mglg% mglg% ACA101108-1 3.15 xx 0.93 100 ACA101108-2 3.12 1.30 108 1.18 100 ACA101108-3 3.73 1.27 127 0.96 100 ACA101108 -4 3.8 xx 0.93 100 S Table 1a) 1 week 70 ° Sample Benzoic acid Oxytocin pH mglg% mglg% ACA101108-1 3.10 xx 0.36 38.7 ACA101108-2 3.07 1.11 93 0 , 47 39.8 ACA101108-3 3.81 1.07 107 0.23 24 ACA101108-4 3.82 xx 0.24 25.8 Table 1b) 2 weeks 70 ° Sample Benzoic acid Oxytocin pH mglg% mglg% ACA101108-1 ND xx 0.29 31.1 ACA101108-2 ND 1.12 112 0.34 28.8 ACA101108-3 ND 1.13 113 0.15 15.6 ACA101108-4 ND xx 0.20 21.5 Table 1c) 535 ÜEVI PS54264EN00 29 Table 2 a) -d): pH, oxytocin and benzoic acid content of the samples in series ACA101123 after up to 3 weeks storage.
    Time 0 Sample Benzoic acid Oxytocin pH mglg% mglg% ACA101123-1 3.25 xx 0.98 100 ACA101123-2 3.22 0.95 95 0.94 100 ACA101123-3 3.25 0.87 87 0.88 100 ACA101123 -4 3.31 xx 0.86 100 Table 2a) 1 week 70 ° Sample Bonzoic acid Oxytocin pH mglg% mglg% ACA101123-1 ND xx 0.75 76.5 ACA101123-2 ND 1.20 120 0.77 81.9 ACA101123-3 ND 1.12 112 0.74 84.1 ACA101123-4 ND xx 0.78 90.7 S Table 2b) 2 weeks 70 ° Sample Benzoic acid Oxytocin pH mglg% mglg% ACA101123-1 3.31 xx 0.52 53.1 ACA101123-2 3.29 1.04 109 0.52 55.3 ACA101123-3 3.25 0.98 112 0.50 56.8 ACA101123-4 3.32 x X 0.50 58.1 Table 2c) 536 091 PS54264SE00 30 3 weeks 70 ° Sample Benzoic acid Oxytocin pH mglg% mglg% ACA101123-1 3.30 xx 0.46 46.9 ACA101123-2 3.25 1.18 124 0.51 54.3 ACA101123-3 3.31 0.82 94.3 0.43 48.9 ACA101123-4 3.33 xx 0.47 54.7 Table 2d) Table 3 a) -c): pH, oxytocin and benzoic acid content in the samples in series ACA101207 after up for 2 weeks storage.
    Time 0 Sample Benzoic acid Oxytocin pH mglg% mglg% ACA101207-1 4.17 xx 0.98 100 ACA101207-Z 3.88 1.06 100 0.95 100 Table 3a) 1 week 70 ° Sample Benzoic acid Oxytocin pH mglg% mglg% ACA101207-1 3.98 xx 0.58 59.2 ACA101207-Z 3.78 1.00 94.3 0.58 61.1 Table 3b) 536 Ü9'l PS54264SEOO 31 2 weeks 70 ° Sample Benzoic acid Oxytocin pH mg / g% mg / g% ACA101207-1 3.75 xx 0.50 51.0 ACA101207-2 3.67 1.06 100.00 0.51 53.7 Table 3c) Table 4. Appearance and viscosity of the samples after up for 3 weeks storage.
    Lagrin ACA101108-1 ACA101108-2 Appearance viscosity Appearance viscosity Temperature Time 70 ° C 2 weeks Clear NA Clear NA Storage ACA1 01108-3 ACA1 01 1 08-4 Appearance viscosity Appearance viscosity Temperature Time 70 ° C 2 weeks Slightly yellow Very low Clear Very low vis vis vis Storage ACA1 01 1 23-1 ACM 01 1 23-2 Appearance viscosity Appearance viscosity Temperature Time 70 ° C 3 weeks Clear NA Clear NA Storage ACA101123-3 ACA101123-4 Appearance viscosity Appearance viscosity Temperature Time 70 ° C 3 weeks Slightly yellow Very low Very Very low viscone Weak yellow viscous 10 15 20 25 30 535 091 PS54264ENO0 32 Storage ACA101207-1 ACA101207-2 Appearance Viscosity Appearance Viscosity Temperature Time 70 ° C 2 weeks Very Low viscose Weak yellow Low viscous. pale yellow Discussion In the previous stability study for oxytocin, the rate of degradation at different pH values was examined. This study showed that the degradation of oxytocin in a buffered aqueous solution of pH 3, stored below 70 ° C for 7 days was about 15% (area%).
    The corresponding degradation at pH 4 was approximately 35% (area%). In Table 1 above, it is shown that degradation of oxytocin in considerably more extensive sub-comparable pH conditions, about 60% and 75%, respectively. These data indicate that the stability of oxytocin may depend not only on the pH but also on the type of acid used for buffering. Furthermore, the data show that Na-CMC in itself has a negative effect on stability.
    Table 2 shows data from analysis of oxytocin solutions in which Na-CMC has been replaced by a neutral cellulose derivative, HPMC, and in which the buffer is 25 mM citrate. These data again show a considerably better stability of oxytocin and the corresponding degradation is within the magnitude of about 20%. Table 3 shows the corresponding degradation of oxytocin in an HPMC-containing formulation in which lactate is used as a buffering agent.
    These data indicate a similar stability of oxytocin in lactate and citrate under comparable pH conditions. Additional formulations in which lactate was used in combination with Na-CMC resulted in precipitation and turbidity of the formulations. This is not seen in the formulations where HPMC is used with lactate, Table 4.
    From Table 4 it can be concluded that formulations with benzoic acid are slightly more yellowish than formulations without this preservative. The lactate-based formulations are slightly more viscous than the citrate-based formulations after storage at 70 ° C.
    The studies presented above also indicate that benzoic acid is compatible with oxytocin and the ingredients used. 535 091 PS54264EN00 33 Conclusions This study shows that the stability of oxytocin in an acidic environment mainly follows the pattern seen previously, ie even better stability is seen at pH 3 than at pH 4.
    It is concluded that oxytocin is more stable in formulations with HPMC as gelling agent than with Na-CMC. The viscosity of the formulations decreases during storage at 70 ° C for both gelling agents. The stability of oxytocin is similar in lactate and citrate under comparable pH conditions. Benzoic acid is also considered a suitable preservative. 10 536 091 PS54264SE00 References 1. Hawe A. et al., Towards heat-stable oxyfocin formulations: Analysis of degradation kinetics and identification of degradation products, Pharmaceutical Research, 2009, 26: 1679-1688. 2. Stability study of oxytocin in aqueous solutions and dry powder, Labagon AB, 2010. 3. Fawcett JP, et al., Binding of parabens to sodium carboxymethylcellulose in oral liquid formulations, Aust. J. Hosp. Pharm., 1996, 26: 552-554. 4. WO / 92/09307 <110> <120> <130> <170> íšïïï <212> <2l3> <220> <22l> <222> <223> <400> 536 Ü91 seq 1ist1ng 2_5T25 SEQUENCE LISTING Pep- Tonic Medica] AB Pharmaceutícaï composition PSS4264SE00 24 Patentïn version 3.5 å PRT _ Homo sap1ens Moo_Res (9) -. (9) AMIDATION 1 âys Tyr I1e G1n êsn Cys Pro Leu G1y <210> <211> <212> <213> <220 > <223> <220> <22l> <222> <223> <220> <22l> <222> <223> <220> <22l> <222> <2Z3> <220> <22l> <222> < 223> <220> <22l> <222> <223> <220> <22l> <222> <223> 2 10 PRT_ _ _ Art1f1c1a1 sequence Oxytocín fragments and variants MISC_FEATURE (1) .. (1) _ X1 can be Cys or noth1ng MISC_FEATURE (2) - (2) _ X2 can be Tyr, Phe or noth1ng MISC_FEATURE (3) .. (3 X3 can ge Iïe, va1, Hoph, Phe, cha or nothing MISC_FEATURE (4) - ( 4), X4 can be G1n, Ser, Thr, C1t, Arg or Daba MISC_FEATURE (7) .. (7) _ _ XS (pos1t1on 7) can be Pro or noth1ng MISC_FEATURE (8) .. (8) '_ X6 (pos1ç1on 8) can be I1e, Leu, noth1ng, va1, Hos, Daba, Thr, Arg or C1t Page 1 <220> <221> <222> <223> <220> <221> <222> <223 > <220> <22l> <222> <223> <400> 536 091 seq ïisting 2_ $ T25 MISC_FEATURE (9)., (9) _ _ X7 (pos1t1on 9) can be G1y, noth1ng or A1a MOD_Res (l0) .. (l0) AMIDATION MISC_FEATURE (10) .. (} 0) X8 Cposwtion 10) can be G1y or nothing 2 åäö Xâä Xaa Xaa ÅSH CYS Xaä Xaâ Xaâ Xåä 5 1 <2l0> <211> <2l2> <2l3> <220> <221> <222> <223> <400> 3 9 PRT Bird Moo_REs (9) - (9) AMIUATION 3 âys Tyr I1e G1n: sn Cys Pro I1e G1y <210> <211> <2l2> < 2l3> <220> <221> <222> <223> <400> 4 9 PRT Fish MOD_RES (9) .. (9) AMIDATION 4 âys Tyr I1e Ser Ésn Cys Pro I1e G1y <2l0> <211> <2l2> <2l3> <220> <221> <222> <223> <400> S 9 PRT _ _ Eisen1a foet1da MOD_RES (9) .. (9) AMIDATION 5 Page 2 536 091 seq 11sting 2_ST2S âys Phe va1 Arg ês Cys Pro Thr G1y <210> <211> <212> <213> <220> <221> <222> <223> <400> 6 9 PRT non mamma1ian vertebrates, feta1 mamma1s MoD_REs (9) .. (9) AMIDATION 6 Cys Tyr I1e G1n êsn Cys Pro Arg G1y 1 <210> <211> <212> <213> <220> <221> <222> <223> <400> 7 9 PRT Homo sapiens MoD_REs (9) - (9) AMIDATION 7 ïys Tyr Phe Gïn is Cys Pro Arg G1y <210> <21l> <212> <213> <220> <223> <220> <221> <222> <223> <400> 8 6 pnn Art1f1c1a1 sequence Oxytocin fragment Mon_Res (6). . (6) AMIDATIQN 8 Cys Tyr I1e G1n êsn Cys 1 <210> <2l1> <212> <213> <220> <223> 9 7 PRT _ Artific1a1 sequence oxytocin fragment Page 3 <220> <22l> <222> <223> <400> Moo_n: s (7) .. (7) AMIoATIoN 9 âys Tyr I1e G1n êsn Cys Pro <210> <2l1> <212> <213> <220> <223> <220> <22l> <222> <223> <400> 10 8 PRT_ _ _ Art1f1c1a1 sequence Oxytocín fragment MoD_REs (8) .. (8) AMIoATIoN 10 äys Tyr Iïe Gïn êsn Cys Pro Leu <210> <211> <212> <213> < 220> <223> <220> <22l> <222> <223> <400> 11 8 PRT Artificia1 sequence Oxytocin fragment MoD_REs (8) .. (8) AMIDATION ll: yr Iïe G1n Asn gys Pro Leu G1y <2lO> <211> <212> <2l3> <220> <223> <220> <22l> <222> <223> <400> 12 7 PRT_ _ _ Art1f1c1a1 sequence oxytocin fragment Moo_Rss (7) .- (7) AM1oAT1oN 12 I1e Gin Asn Cys Pro Leu G1y 536 09¶ seq 1isting 2_ST25 Page 4 1 <210> <211> <212> <213> <220> <223> <220> <221> <222> <223> <400> 536 091 seq ïisting 2_ST25 13 6 PRT_ Art1fícia1 sequence oxytocín fragment Moo_REs (6) .. (6) AMIDATION 13 ïïn Asn Cys Pro šeu G1y <210> <2l1> <212> <213> <220> <223> <220> <221> <222> <223> <400> 14 5 PRT_ _ _ Art1f1c1a1 sequence Oxytocin fragment MOD_RES (S) - (5) AM1oAT1oN 14 Iïe G1n Asn Cys Pro 1 5 <210> <211> <212> <213 > <220> <223> <220> <221> <222> <223> <400> 15 10 nnn Art1f1c1a1 sequence Oxytocin variant MOD_RES (10) .. (10) AMIDATION 15 Cys Tyr I1e G1n Asn Cys Pro Leu G1y G1y 1 5 10 <210> <211> <212> <213> 16 6 PRT Artificíaï sequence Page 5 <220> <223> <220> <221> <222> <223> <400> 536 091 seq ïistíng 2_ST25 oxytocin variant MoD_REs (6) .. (6) AMIDATION 16 ï1n Asn Cys Pro äeu Leu <2l0> <2l1> <212> <213> <220> <223> <220> <221> <222> <223> <400> 17 9 PRT Artificiaï sequence Oxytocin variant Moo_nEs (9) .. (9) AMIDATION 17 (lïys Tyr va1 Thr Ašsn Cys Pro Leu Gïy <2l0> <2l1> <212> <213> <220> <223> <220> < 221> <222> <223> <220> <221> <222> <223> <400> 18 9 PRT Artificiaï sequence Oxyt ocín variant MISC_FEATURE (3) .. (3) X is Hoph MOD_RES (3) .. (3) AMIDATION 18 âys Tyr Xaa Thr êsn Cys Pro va1 G1y <2l0> <21l> <212> <213> <220> < 223> 19 9 PRT Artifícía1 sequence Oxytocin variant Page 6 <220> <22l> <222> <223> <220> <22l> <222> <223> <400> 536 091 seq ïísting 2_ST25 MISC_FEATURE (42 - (4 ) X 1s C1t MOD_RES (9) - (9) AMIDATION 19 âys Tyr Phe Xaa êsn Cys Pro Leu G1y <210> <211> <2l2> <2l3> <220> <223> <220> <22l> <222 > <223> <220> <22l> <222> <223> <220> <22l> <222> <223> <400> 20 9 PRT_ _ _ Art1f1c1a1 sequence Oxytocín variant MISC_FEATURE (3) .. (3) X 1s cha MISC_FEATURE (8) .. (8) X 1s Hos MOD_REs (9) .. (9) AMIDATION 20 âys Tyr Xaa Arg êsn Cys Pro Xaa A1a <210> <211> <2l2> <213> <220> < 223> <220> <22l> <222> <223> <220> <22l> <222> <223> <220> <22l> 21 9 PRT Artifíc1a1 sequence Oxytocin variant M1sC_FEATuRa (4) .. (4) X 1s Daba MISC_FEATURE (82 - (8) X 1s Daba MOD_RES Page 7 <222> <223> <400> 536 091 seq Tisting 2_ST25 (9) .- (9) AMI oAuoN 21 Cys Tyr va1 Xaa Asn cys Pro Xaa A1a 1 S < 210> <2ll> <212> <213> <220> <223> <220> <22l> <222> <223> <220> <22l> <222> <223> <220> <22l> <222> <223> <220> <22l> <222> <223> <400> 22 9 nuc Art1f1c1a1 sequence Oxytocin variant M1sC_FEATuRE (s). .
    X 1s Hoph MISC_FEATURE (4) .. (4) X 1s Daba MISC_FEATURE (8) - (S) X 15 C1t MOD_RES (9) .. (9) AMIDATION 22 âys Tyr Xaa Xaa êsn Cys Pro Xaa A1a <210> <211> <212> <213> <220> <223> <220> <22l> <222> <223> <400> 23 9 PRT Artificiaï sequence Oxytocin variant MOD_RES (9) .. (9) AMIDATION 23 âys Tyr Phe Arg êsn Cys Pro vaï A1a <2l0> <2ll> <212> 24 9 PRT Art1ficia1 sequence Page 8 <220> <223> <220> <22l> <222> <223> <220> <22l> <222> <223> <220> <22l> <222> <223> <400> Oxytocin variant MISC_FEATuRE (3) .. (3) X 1s cha MIscJeATuRs (4) .. (4) x 1 s c1 t MOD_RES (9) .. (9) AMIDATION 24 536 091 seq ïistíng 2_ST25 âys Tyr Xaa Xaa êsn Cys Pro Arg Gïy Page 9 <110> <120> <130> <160> <170> <210> <2l1> <2l2> <2l3> <220> <221> <222> <223> <400> 535 Ü91 seq ïisting 2_ST25 translated to11 SE SEQUENCE LIST Pep-Tonic Medicai AB Pharmaceutical Composition PS54264SE00 24 Patentln version 3.5 1 9 PKT _ Homo sapiens MOD_RES (9) - (9 ) AMIDERING 1 âys Tyr Iie Gin êsn cys Pro Leu G1y <2l0> <211> <2l2> <2l3> <220> <223> <220> <221> <222> <223> <220> <221> <222> <223> <220> <221> <222> <223> <220> <22l> <222> <223> <220> <221> <222 > <223> <220> <221> <222> <223> 2 10 PRT Artificie11 sequence Oxytocin fragments and variants MIsc_FEATuRE (1) - (1) _ _ X1 can be Cys e11er nothing MISC_FEATURE (2) .. (2) _ _ X2 can be Tyr, Phe e11er nothing MISC_FEATURE. . (s) _ _ X3 can be I1e, va1, Hoph, Phe, Cha eiier nothing MISC_FEATURE (4) - (4) _ X4 can be Gin, ser, Thr, cit, Arg e11er Daba MISC_FEATURE (7) .. ( 7) _ _ _ X5 (position 7) can be Pro e11er nothing MIsc_FEATuRe (8) .. (8) X6 (position 8) can be Iie, Leu, nothing, vai, Hos, Daba, Thr, Arg eïïer Cit Page 1 <220> <221> <222> <223> <220> <221> <222> <223> <220> <221> <222> <223> <400> Xaa Xaa Xaa Xaa Asn Cys Xaa Xaa Xaa Xaa 1 5 <210> <211> <212> <2l3> <220> <221> <222> <223> <400> 536 091 seq ïísting 2_ST25 translated to11 SE MISC_FEATURE (9> - C92...
    X7 (pos1t1on 9) kan vara G1y, 1ngent1ng e11er Aïa Moo_REs (10) .. (10) Amzosaxnc MIsc_FEATuRE (10) .. (; 0) _ _ X8 (pos1tion 10) kan vara G1y e11er 1ngent1ng 2 10 3 9 PRT Fàgeï MoD_REs (9) .. (9) AMIDERING 3 Cys Tyr Iïe Gïn êsn Cys Pro Iïe Gïy 1 <210> <211> <212> <213> <220> <221> <222> <223> <400> 4 9 PRT Fisk MoD_REs (9) - (9) AMIDERING 4 âys Tyr I1e Ser êsn Cys Pro I1e G1y <210> <211> <212> <2l3> <220> <221> <222> <223> <400> S 9 PRT _ Eísenia foet1da MOD_RES (9) .. (9) AMIDERING 5 Page 2 <2l0> <2l1> <2l2> <213> <220> <22l> <222> <223> <400> 536 OBW seq ïisting 2_ST25 translated ti11 SE âys Phe vaï Arg êsn Cys Pro Thr G1y 6 9 PRT non-mammaïie vertebrates, feta1a mamaïier MOD_RES (9) - (9) AMIDATION 6 gys Tyr I1e G1n êsn Cys Pro Arg G1y <2l0> <211> <2l2 > <213> <220> <22l> <222> <223> <400> 7 9 PRT _ Homo sap1ens Moo_Res (9) .. (9) AMIDERING 7 gys Tyr Phe G1n êsn Cys Pro Arg G1y <2l0> <2l1 > <2l2> <2l3> <220> <223> <220> <22l> <222> <223> <400> 8 6 PRT Artif1cie11 sequence Oxytocin fragment MOD_RES (6 ) .. (6) AMIDERING 8 Cys Tyr I1e G1n êsn Cys 1 <2l0> <211> <2l2> <2l3> <220> <223> 9 7 PRT Artificie11 sequence oxytocin fragment Page 3 535 G91 seq ïisting 2_ST2S translated ti11 SE < 220> <22l> MOD_RES <222> (7) .. (7) <223> AMIDERING <400> 9 šys Tyr I1e G1n êsn cys Pro <2l0> 10 <211> 8 <212> PRT <213> Art1fícíe11 sekvens < 220> _ <223> 0xytoc1nfragment <220> <22l> MOD_REs <222> (8) .. (8) <223> AMIDERING <400> 10 âys Tyr I1e G1n êsn cys Pro Leu <2l0> 11 <211> 8 < 212> PRT <213> Artificie11 sequence <220> <223> Oxytocin fragment <220> <22l> MOD_RES <222> (8) .. (8) <223> AMIDERATION <400> 11 Tyr Iïe G1n Asn gys Pro Leu G1y 1 <2l0> 12 <21l> 7 <212> PRT_ _ _ <213> Art1f1c1e11 sequence <220> <223> Oxytocin fragment <220> <22l> MOD_RES <222> (7) .. (7) <223> AMIDERING <400 > 12 I1e G1n Asn Cys Pro Leu G1y Page 4 536 091 seq ïisting 2_ST2S translated ti11 SE <2lO> 13 <211> 6 <2l2> PRT <213> Artificieïï sequence <220> <223> Oxytocin fragment <220> <221> MOD_RES <222> (6) .. (6) <223> AMIDERING <400> 13 ïïn Asn Cys Pro šeu G1y <2lO> 14 <211> 5 <2l2> PRT_ <213> Art1fície11 sequence <220> <223> Oxytocin fragment <220> <221> MOD_RES <222> (5) .. (5) <223> AMIDERING <400 > 14 I1e G1n Asn Cys Pro 1 S <2lO> 15 <21l> 10 <2l2> PRT <2l3> Artifícieïï sequence <220> <223> Oxytocinvariant <220> <221> fl OD_RES <222> (10) .. (10) .. ) <223> AMIDERING <400> 15 Cys Tyr I1e Gïn Asn Cys Pro Leu G1y G1y 1 S 10 <2lO> 16 <2l1> 6 <2l2> PRT <213> Artificie11 sequence Page 5 536 091 seq ïisting 2_ST2S translated tí11 SE < 220> <223> Oxytocinvariant <220> <221> MOD_RES <222> (6) .. (6) <223> ÅMIDERING <400> 16 É1n Asn Cys Pro Leu Leu 5 <210> 17 <211> 9 <212> PRT <213> Artificial11 sequence <220> <223> Oxytocin variant <220> <221> MOD_RES <222> (9) .. (9) <223> AMIDERING <400> 17 âys Tyr Va1 Thr êsn Cys Pro Leu G1y <210 > 18 <211> 9 <212> PRT <2l3> Artifície11 sequence <220> _ <223> 0xytocinvar1ant <220> <22l> MISC_FEATURE <222> (3) .. (3) <223> X is Hoph <220> <221> MOD_RES <222> (3) .. (3) <223> AMIDERING <400> 18 Cys Tyr Xaa Thr êsn Cys Pro va1 G1y 1 <2 10> 19 <211> 9 <212> PRT <213> Artificial11 sequence <220> <223> Oxytocin variant Page 6 536 091 seq ïisting 2_ST25 translated to11 SE <220> <221> MISC_FEÅTURE <222> (4) .. (4) ) <223> X is Cit <220> <221> MOD_RES <222> (9) .. (9) <223> AMIDERING <400> 19 Eys Tyr Phe xaa êsn Cys Pro Leu G1y <210> 20 <2l1> 9 <212> PRT <213> Artificieïsequence <220> <223> Oxytocin variant <220> <22l> MI $ C_FEÅTURE <222> (3) .. (3) <223> X är Cha <220> <221> MISC_FEATURE < 222> (8) .. (8) <223> X är Hos <220> <221> MOD_RES <222> (9) .. (9) <223> AMIDERING <400> 20 âys Tyr xaa Arg êsn Cys Pro xaa A1a <210> 21 <211> 9 <212> PRT <213> Artificieïï sequence <220> _ _ <223> 0xytoc1nvar1ant <220> <22l> MISC_FEÅTURE <222> (4) .. (4) <223> X är Daba <220> <221> MISC_FEATURE <222> (8) .. (8) <223> X är Daba <220> <221> MOD_RES Page 7 535 091 seq ïistíng 2_ST25 translated to11 SE <222> (9) .. (9) <223> AMIDERING <400> 21 âys Tyr Va1 Xaa is Cys Pro Xaa A1a <2l0> 22 <211> 9 <2l2> PRT <213> Art1fície11 sequence <220> <223> oxytocin variant <220> < 221> MISC_FEATURE <222> (3) .. (3) <223> X is Hoph <220> <221> MIsc_FEATuRE <222> (4) .. (4) <223> X is Daba <220> <221> MISC_FEATURE <222> (8) .. (§) <223> x är C1t <220> <221> MOD_RES <222> (9) .. (9) <223> AMIDERING <400> 22 âys Tyr Xaa Xaa êsn Cys Pro Xaa A1a <2l0> 23 <211> 9 <2l2> PRT »<213> Artificieïï sequence <220> _ <223> 0xytocinvar1ant <220> <221> MOD_RES <222> (9) .. (9) <223> AMIDERING <400> 23 âys Tyr Phe Arg êsn Cys Pro va1 A1a <2l0> 24 <21l> 9 <2l2> PRT <213> Artíficie11 sequence Page 8 536 ÜÉVI seq 1isting 2_ST2S translated to11 SE <220> <223> Oxytocin variant <220 > <22l> MISC_FEATURE <222> (3) .. (3) <223> X är Cha <220> <221> MISC_FEATURE <222> (4) .. (4) <223> X är Cít <220> < 22l> MOD_RES <222> (9) .. (9) <223> AMIDERING <400> 24 Cys Tyr Xaa Xaa êsn Cys Pro Arg Gïy 1 Page 9
    权利要求:
    Claims (9)
    [1] 1. A pharmaceutical composition comprising a. oxytocin (SEQ ID NO:1), and/or one or more fragment(s) and/or variant(s)thereof according to SEQ ID NO:2, as well as pharmaceutically acceptablesalts thereof; and b. at least one non-ionic cellulose ether; wherein SEQ ID NO:2 is X1-X2-X3-X4-Asn-Cys-X5-X6-X7-X8-NH2 wherein X1 is selected from the group consisting of Cys and nothing; X2 is selected from the group consisting of Tyr, Phe, and nothing; X3is selected from the group consisting of lle, Val, Hoph, Phe, Cha, andnothing; X4 is selected from the group consisting of Gln, Ser, Thr, Cit, Arg, and Daba;X5 is selected from the group consisting of Pro and nothing; X6 is selected from the group consisting of lle, Leu, nothing, Val, Hos, Daba,Thr, Arg, and Cit; X7 is selected from the group consisting of Gly, nothing, and Ala; X8 is selected from the group consisting of Gly and nothing; wherein said pharmaceutical composition has a pH within the range of betweenabout 3 and about 4. _ A pharmaceutical composition according to claim 1, wherein said one or more fragment(s) and/or variant(s) of oxytocin is/are selected from the group consistingof the peptides corresponding to SEQ ID NO:3-SEQ lD NO:24. . A pharmaceutical composition according to claim 1 or 2, wherein said pH is within the range of between about 3 and about 3.8, such as between about 3 and about3.5, or about 3 and 3.3. . A pharmaceutical composition according to any of claims 1-3, wherein the pH of said composition is regulated by adding a pH regulating agent to said composition. . A pharmaceutical composition according to claim 4, wherein said pH regulating agent is a buffer. . A pharmaceutical composition according to claim 5, wherein said buffer is a lactate or a citrate buffer. 7. 10. 11. 1
    [2] 2. 1
    [3] 3. 1
    [4] 4. 1
    [5] 5. 1
    [6] 6. 1
    [7] 7. 1
    [8] 8. A pharmaceutical composition according to any of claims 1-6, further comprising apreservative, such as benzoic acid. A pharmaceutical composition according to any of the preceding claims, whereinsaid at least one non-ionic cellulose ether is selected from the group consisting ofmethyl cellulose (MC), hydroxypropylmethylcellulose (HPMC),hydroxypropylcellulose (HPC), hydroxypropylmethylcellulose (HPMC),hydroxyethylethylcellulose (HEEC) and hydroxyethylmethyl cellulose (HEMC). A pharmaceutical composition according to any of claims 1-7, wherein said at leastone non-ionic cellulose ether is hydroxypropylmethylcellulose (HPMC). A pharmaceutical composition according to any of claims 1-7, comprising oxytocin(SEQ ID NO:1) and hydroxypropylmethylcellulose (HPMC). A pharmaceutical composition according to any of the preceding claims, whereinthe concentration of oxytocin and/or one or more fragment(s) and/or variant(s)thereof as defined in claim 1 is about 1 mg/g of said pharmaceutical composition.A pharmaceutical composition according to any of claims 1 or 3, said compositioncomprising about: 1 mg/g oxytocin, 2 wt% hydroxypropyl methylcellulose in 25 mM citrate buffer, and optionally 1 mg/g benzoic acid. A pharmaceutical composition according to any of claims 1 or 3, said compositioncomprising about: 1 mg/g oxytocin, 2 wt% hydroxypropyl methylcellulose in 25 mM lactate buffer, and optionally 1 mg/g benzoic acid. A pharmaceutical composition according to any of the preceding claims for use asa medicament. A pharmaceutical composition according to any of the preceding claims for topicaluse. A pharmaceutical composition according to any of the preceding claims for vaginaluse. A pharmaceutical composition according to any of the preceding claims for use inthe treatment of a climacteric disorder, such as vaginal atrophy. Use of at least one non-ionic cellulose ether for improving the stability of apharmaceutical composition comprising oxytocin (SEQ ID NOI1) and/or one ormore fragment(s) and/or variant(s) of oxytocin, as defined in claim 1 (SEQ lDNO:2) or claim 2. 1
    [9] 9. 20. 21. 22. 23. 24. 36 Use according to claim 18, wherein said at least one non-ionic cellulose ether ishydroxypropylmethylcellulose (HPMC).Use according to claim 18, wherein said pharmaceuticai composition is as definedin any of claims 1-13.Method for preparing a pharmaceuticai composition with improved stabilitycomprising the steps of:a) preparing a buffer solution and adjusting the pH thereof,b) adding oxytocin and/or one or more fragment(s) and/or variant(s) thereofas defined in claim 1, to said buffer solution, and thereafterc) adding at least one non-ionic cellulose ether to the solution obtained instep b);wherein optionally a preservative, such as benzoic acid, is added to thebuffer solution of step a) before said oxytocin and/or one or more fragment(s)and/or variant(s) thereof is/are added to said buffer solution,and wherein the pH of said pharmaceuticai composition is regulated to fallwithin the range of between about 3 and about 4, such as within the range ofabout 3 and about 3.5.Method according to claim 21, for the preparation of a pharmaceuticai compositionaccording to any of claims 1-13.l/lethod according to any of claims 21-22, wherein said at least one non-ioniccellulose ether is hydroxypropylmethylcellulose (HPMC).A pharmaceuticai composition comprisinga. oxytocin (SEQ ID NO:1), and/or one or more fragment(s) and/or variant(s)thereof according to SEQ lD NO:2, as well as pharmaceutically acceptablesalts thereof; andb. at least one non-ionic cellulose ether;wherein SEQ ID NO:2 isX1-X2-X3-X4-Asn-Cys-X5-X6-X7-X8-NH2whereinX1 is selected from the group consisting of Cys and nothing,X2 is selected from the group consisting of Tyr, Phe, and nothing,Xßis selected from the group consisting of Ile, Val, Hoph, Phe, Cha, and nothing,X4 is selected from the group consisting of Gln, Ser, Thr, Cit, Arg, and Daba,X5 is selected from the group consisting of Pro, and nothing,X6 is selected from the group consisting of Ile, Leu, nothing, Val, Hos, Daba, Thr,Arg, and Cit, 25. 37 X7 is selected from the group consisting of Gly, nothing, and Ala, X8 is selected from the group consisting of Gly, and nothing, wherein oxytocin and/or one or more fragment(s) and/or variant(s) thereofaccording to SEQ ID NO:2, or a pharmaceuticaily acceptable sait thereof is/arepresent in said pharmaceutical composition in a concentration of about 0.5 toabout 1.5 mg/g of the totai pharmaceutical composition, and said at least one non-ionic celiulose ether is present in an amount of about 1-3 wt%, such as about 2wt%, in a pH regulating agent, such as a buffer, said pH regulating agent having a concentration of about 25 mM to about 100 mM. A pharmaceutical composition according to ciaim 24, wherein said at ieast onenon-ionic cellulose ether is hydroxypropyi methylcellulose and is present in anamount of about 2 wt% in a buffer with a concentration of about 25 mM, said buffer being a lactate or a citrate buffer.
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    同族专利:
    公开号 | 公开日
    EP2696882B1|2015-03-04|
    BR112013026153A2|2019-09-17|
    SE536091E5|2015-04-21|
    JP2014515747A|2014-07-03|
    CN103596582A|2014-02-19|
    US20140171369A1|2014-06-19|
    KR101940341B1|2019-01-18|
    US9034821B2|2015-05-19|
    KR20140019830A|2014-02-17|
    CN103596582B|2016-03-16|
    RU2013147739A|2015-05-20|
    SG194074A1|2013-11-29|
    CA2832664C|2020-03-24|
    IL228821A|2016-08-31|
    ES2535521T3|2015-05-12|
    AU2012241805A1|2013-10-24|
    IL228821D0|2013-12-31|
    EP2696882A1|2014-02-19|
    RU2605288C2|2016-12-20|
    AU2012241805B2|2016-08-11|
    ZA201308157B|2015-01-28|
    WO2012140216A1|2012-10-18|
    PL2696882T3|2015-08-31|
    SE1150324A1|2012-10-15|
    CA2832664A1|2012-10-18|
    HK1189158A1|2014-05-30|
    DK2696882T3|2015-04-27|
    JP5993442B2|2016-09-14|
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    法律状态:
    2015-04-21| LMSE| Limitation by swedish patent and registration office|Effective date: 20141001 |
    2015-12-15| NUG| Patent has lapsed|
    优先权:
    申请号 | 申请日 | 专利标题
    SE1150324A|SE536091E5|2011-04-14|2011-04-14|SE1150324A| SE536091E5|2011-04-14|2011-04-14|
    DK12715932T| DK2696882T3|2011-04-14|2012-04-13|Pharmaceutical composition|
    RU2013147739/15A| RU2605288C2|2011-04-14|2012-04-13|Pharmaceutical composition|
    ES12715932.5T| ES2535521T3|2011-04-14|2012-04-13|Pharmaceutical composition|
    JP2014504346A| JP5993442B2|2011-04-14|2012-04-13|Pharmaceutical composition|
    CA2832664A| CA2832664C|2011-04-14|2012-04-13|Pharmaceutical composition|
    AU2012241805A| AU2012241805B2|2011-04-14|2012-04-13|Pharmaceutical composition|
    SG2013074133A| SG194074A1|2011-04-14|2012-04-13|Pharmaceutical composition|
    CN201280027310.1A| CN103596582B|2011-04-14|2012-04-13|Pharmaceutical composition|
    PL12715932T| PL2696882T3|2011-04-14|2012-04-13|Pharmaceutical composition|
    US14/111,072| US9034821B2|2011-04-14|2012-04-13|Pharmaceutical composition|
    BR112013026153A| BR112013026153A2|2011-04-14|2012-04-13|PHARMACEUTICAL COMPOSITION; USE OF AT LEAST ONE NON-IONIC CELLULOSE ETHER; METHOD FOR PREPARING A PHARMACEUTICAL COMPOSITION; METHOD FOR TREATING AND / OR PREVENTING A CLIMATE CLIMATE; USE OF A PHARMACEUTICAL COMPOSITION; AND KIT|
    PCT/EP2012/056813| WO2012140216A1|2011-04-14|2012-04-13|Pharmaceutical composition|
    EP12715932.5A| EP2696882B1|2011-04-14|2012-04-13|Pharmaceutical composition|
    KR1020137029608A| KR101940341B1|2011-04-14|2012-04-13|Pharmaceutical composition|
    IL228821A| IL228821A|2011-04-14|2013-10-10|Pharmaceutical composition comprising oxytocin|
    ZA2013/08157A| ZA201308157B|2011-04-14|2013-10-31|Pharmaceutical composition|
    HK14102115.0A| HK1189158A1|2011-04-14|2014-03-03|Pharmaceutical composition|
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