FR2511866A1 - PHARMACEUTICAL FORMULATIONS OF HUMAN PROINSULIN - Google Patents

Abstract

THE INVENTION CONCERNS A PHARMACEUTICAL COMPOSITION COMPRISING HUMAN PROINSULIN IN ASSOCIATION WITH A PHARMACEUTICALLY ACCEPTABLE SUPPORT, THIS COMPOSITION BEING USEFUL FOR CONTROL OF A DIABETIC STATE AND FOR PROMOTING THE OBTAINING OF NATURAL HORMONAL HOMEOSTASE, IN ORDINARY MITIGATE OR SENSITIVELY BRAKE DIABETIC COMPLICATIONS.

Description

PHARMACEUTICAL FORMULATIONS OF HUMAN PRO-INSULIN.

  Diabetes mellitus is a metabolic disorder characterized by the fact that body tissues fail to oxidize carbohydrates on a diet

  normal Its most important factor is a defi-

  insulin science Over the past 60 years,

  people with diabetes have been considered

  aided by receiving regulated amounts of

  sulin Currently, the insulin used by diabetics has been isolated from the pancreas of animals, usually from cattle and pigs. Insulin from cattle and pigs has a different structure.

  that of insulin produced by the human pancreas.

  Recently, using the DNA methodology (DNA = acid

  deoxyribonucleic) recombinant, it became pos-

  sible to prepare insulin identical to that produced by the human pancreas The use of this insulin will allow diabetics to imitate the

  natural system more tightly than it was pos-

sible so far.

  Nevertheless, it has long been recognized that the mere administration of insulin to a diabetic is insufficient to restore and / or maintain the normal metabolic state. Although linsulin manifests its effect on the metabolism of carbohydrates, diabetes mellitus causes additional disorders. most, if not all, of which relate to the structure and function of blood vessels

  leading to these disorders are rarely corrected com-

  completely by conventional insulin therapy.

  The vascular anomalies associated with the dia-

  are stupid, called "complications of diabetes".

  Eltes coiisi sttnt generally in micro- changes

  angiopathics giving rise to damage to the retina

  and kidney Neuropathy is a complica-

  additional tliabetic action which may be related

Z

251 1866

  Lion or Elol, diir'e ellellmt Ott yes (lil'ectllemmt; ave'c les changesls mi icro-angiopahitli (luts p Gécités -Par mli les

  specific manifestations (complications of the dia-

  animal, there are, for example: (l) The diseases of ú 10 cilj in particular the retinopathy, the formation of the cataract,

  1 e g'lauconie el le-s paal; iysies l II Lts Usuair Irs extra ocutilai-

  res; (2) diseases of the mouth, especially gingivitis, increased occurrence of dental caries, periodontitis and a stronger resorption of the alveolar bone; (3) motor, sensory and autonomous neuropathies; (4) large vessel disease; (5) micro-angiopathy; (6) diseases of the

  skin, especially xantlioma (diabetes mellitus, necrosis

  lipid biosis of diabetes mellitus, furunculosis, yeast infection and pruritus; (7) kidney disease,

  including diabetic glomerulosclerosis, nephro-

  arteriolar sclerosis and pyelonephritis; and (8) problems during pregnancy, including an increase in the birth of large babies, birth of stillbirths, miscarriages, '

  neonatal deaths and birth defects.

  Many and perhaps all of the diabetic complications result from the fact that insulin alone cannot bring the body back to normal.

its natural hormonal balance.

  The present invention relates to comiposi-

  Pharmaceutical measures to achieve and maintain, in a diabetic subject, a state closer to natural th 6 m hormone deostasis than that which can be obtained by the administration of insulin alone. Therefore, the present invention relates to a

  pharmaceutical composition comprising pro-insulin

  human line in association with pharmaceutical support

ceutically acceptable.

  The essential con Isti t Luant of the compositions

  pharmaceutical of the present invention is the pro-

  human insulin Administration of human pro-insulin using a composition according to the present invention

  tion will allow more natural use

  glucose and better glycemic control

  crumb, thereby alleviating diabetic complications

  harmful that have been described above.

  Human proinsulin can be obtained in various ways, in particular by organic synthesis, by isolation of the human pancreas according to the conventional methodology and, more recently, according to the methodology.

  with recombinant deoxyribonucleic acid.

  Broadly speaking, the production of pro-insulin by adopting the recombinant deoxyribonucleic acid methodology consists in obtaining, by isolation and / or by construction, a sequence

  for coding deoxyribonucleic acid for se-

  quence of amino acids of human -pro-insulin.

  Then the deoxyribonucleic acid of

  human proinsulin in the reading phase in a

  appropriate vehicle for cloning and expression

  hicule is used to transform a micro-organ

  appropriate to me, after which the transformed micro-organism is subjected to fermentation conditions leading to: (a) the production of additional copies of the

  vector containing the pro-insulin gene and (b) ex-

  pressure of pro-insulin or a precursor

pro-insulin.

  If the expression product is a precursor

  proinsulin sister, it will generally include the amino acid sequence of human proinsulin joined, at its amino terminus, to a fragment of a protein normally expressed in the gene sequence in which the proinsulin gene has summer

  introduces the amino acid sequence of the pro-

  insulin is joined to the protein fragment by ltin-

  through a specifically cleavable seat, no-

  such as methionine This product is usually

  called "fused gene product".

  The amino acid sequence of the pro-

  insulin is cleaved from the fused gene product using cyanogen bromide, after which the sulfhydryl fractions of cysteine from the amino acid sequence of pro-insuliin are stabilized

  by transformation into their corresponding S-sulfonates

  dants. We purify pro-insulin S-sulfonate

  obtained, then transform the pro- S-sulfonate

  insulin purified into pro-insulin by formation of

  three correctly located disulfide bonds.

  The proinsulin obtained is then purified.

  As indicated, the compositions of the present invention are useful in promoting the production of natural hormonal homeostasis and

  in this way prevent or slow down or slow down significantly-

  the well-known diabetic complications that

have been mentioned above.

  It is recognized that some diabetics are unable to receive insulin effectively by subcutaneous injection due to the presence, at the site of the injection, of proteases which destroy

  insulin quickly before it can be ab-

  absorbed into the blood stream and transported to the receptor seats If these diabetics must absolutely receive insulin, they must receive it by intravenous injection.

  repeated and essential intravenous drugs are not

  portunes due to the harmful effect they exert

  hundred on the recipient's veins with infections

  q (associated with it We discovered that pro-in-

  Human suli li was not degraded by these insulin degrading Caases which, therefore, could be adhered by subcutaneous injection Its sta-

  bility and, therefore, its availability favor ob-

  tention of a natural hormonal homeostasis.

  Following recent studies l Podlecki et al., "Diabetes", 31, supplement 2, 126 A (1982) l, we

  also noted that human proinsulin is

  killed inside d (target tissue, for example,

  fat cells Although its intra-

  cellular at a molecular level is still undetermined, these discoveries support

  further the description given in this specification

  cification that human proinsulin

  plays an active role and is essential for obtaining

  tion of natural hormonal homneostasis.

  In "Diabetes" 31, supplement 2, 135 A

  (1982), Schluter et al describe demon-

  trant that the binding of the human insulin receptor is favored pal lia pleselce (o de p'o-insulin Ihuma III i n (,

  These results also further support the description

  tion of this specification that the

  availability and presence of human pro-insulin

  do not promote or restore natural hormonal hoteostasis. Of course, the amount in which the compositions of the present invention are to be

  used to maintain natural hormonal homeostasis

  or to achieve a state closer to natural hormonal homeostasis in a diabetic subject, will depend on the severity of (the condition

  diabetic In addition, this amount will vary depending on

  Finally, the amount of

  composition administered and the frequency of this ad-

  ministration will be left: s the choice of tédlecin par-

  However, as a general rule, based on the fact that 1 mg of human pro-insulin gives about 3.5 units of human insulin activity, the dosage of human pro-insulin will be in the interval giving about 0.02 to about 5 units of human insulin activity per kg d (per body weight per day, preferably about 0.1 to about 1 unit of human insulin activity per kg of weight

of the body per day.

  The composition is administered by the

  rentérale, notanuinent, subcutaneously,

  intramuscular and intravenous

  positions of the present invention include 11 in-

  active ingredient, namely human proinsulin

  together with a pharmaceutically acceptable carrier

  ble for the latter and possibly with dlau-

  very therapeutic ingredients The total amount of the active ingredient present in the composition is

  is between about 99.99 and about 0.01% by weight.

  The medium must be acceptable in the sense that it must

  be compatible with other components of the

  sition and it must not be harmful to the recipient

of it.

  The compositions of the present invention which are suitable for administration by the

  parenteral, advantageously include suspen-

  sterile aqueous solutions and / or solutions

  pharmaceutically active ingredients, these solutions

  or suspensions being, preferably, made isotoni-

  with the recipient's blood using general-

  sodium chloride, glycerin, glu-

  cose, mannitol, sorbitol and known analogues In addition, the compositions may contain one or 1 other adjuvant L such as buffering agents, preservatives, dispersing agents, agents promoting rapid onset of activity, agents promoting prolongation of activity and other agents known As specific preservatives, there will be mentioned, for example, phenol,

  m-cresol, methyl p-hydroxybenzoate and others.

  As specific buffering agents, mention will be made, for example, of sodium phosphate, sodium acetate,

sodium citrate and others.

  In addition, an acid such as

  l 1 hydrochloric acid, or a base such as hydro-

  Sodium xyde to adjust the fold As a rule,

  the fold of the aqueous composition will be between

  about 2 and about 8, preferably between about

6.8 and around 8.

  Among other suitable additives are

  tionnelra, for example, Itionl zinc bi vallelt (the amount of which, if present, is generally between about 0.1 mg and about 3 mg per 100 units of human proinsulin, as well as a protamine salt (per example, in the form of its sulfate) the amount of which, if present, is generally between approximately 0.5 mg and approximately 20 mg per 100 units

of human pro-insulin.

  Examples of compounds will be given below.

  particular pharmaceutical according to the pre-

invention.

Example 1

  Formulation of neutral regular human proinsulin l 40 units of human proinsulin per 3 ECm. To prepare 10 cm 3 of the composition, the following are mixed: Human proinsulin (3.5 units / mg) 114 mg (400 units) Phenol (distilled) 20 mg Glycerin 160 mg Water and either 10% hydrochloric acid or 10% sodium hydroxide: sufficient quantity to obtain a composition with a volume of 10 cm 3 and

a final p H of 7-7.8.

Example 2

  Protamine-zinc human pro-insulin formulation

  l 40 units of human pro-insulin per cm 3 l.

  To prepare 10 cm 3 of the composition, the following are mixed: Human pro-insulin (3.5 units / mg) 114 mg (400 units) Phenol (distilled) 25 mg Zinc oxide 0.95-3.8 mg Glycerin 160 mg Protamine sulfate 32-64 mg Sodium phosphate (crystals) 38 mg Water and either 10% hydrochloric acid or 10% sodium hydroxide 'sufficient to obtain a composition with a volume of 10 cm 3 and

a final fold of 7.1-7.4.

Example 3

  Isophane-protamine human pro-insulin formulation

  l 40 units of human pro-insulin per cm 3 l.

  To prepare 10 cm 3 of the composition, one ml: l alige: Human pro-insulin (3.5 units / mg) 114 mg (400 units) m-cresol (distilled) 16 mg Phenol (distilled) 6.5 mg Glycerin 160 mg Protamine sulphate 9.6-19.2 mg Sodium phosphate (crystals) 38 mg Water and either 10% hydrochloric acid or 10% sodium hydroxide o: sufficient quantity to obtain a composition of a volume of 10 cin 3 and

from a final fold of 7.1-7.4.

Example 4

  Formulation of human pro-insulin zinc l 40 units of human pro-insulin per cm 3 l, To prepare 10 cm 3 of the composition, the following are mixed: Human pro-insulin (3.5 units / mg) 114 mg (400 units) Sodium acetate (anhydrous) 16 mg Sodium chloride (granular) 70 mg m, ethyl phydroxybenzoate 10 mg Zinc oxide 1-8 mg Water and either 10% hydrochloric acid or sodium hydroxide at 10%: sufficient quantity to obtain a composition with a volume of 10 cm 3 and

a final p H of 7.2-7.5.

2511866

Claims (3)

CLAIMS 2511866   1. Pharmaceutical composition comprising pro-insulin   human in combination with a pharmaceutically acceptable carrier.   2. Composition according to claim 1, characterized in   what it contains the bivalent zinc ion.   3 Composition according to claim 1, characterized in   that it contains a protamine salt.