CA1173388A - Analogues of insulin - Google Patents
Analogues of insulinInfo
- Publication number
- CA1173388A CA1173388A CA000385067A CA385067A CA1173388A CA 1173388 A CA1173388 A CA 1173388A CA 000385067 A CA000385067 A CA 000385067A CA 385067 A CA385067 A CA 385067A CA 1173388 A CA1173388 A CA 1173388A
- Authority
- CA
- Canada
- Prior art keywords
- insulin
- des
- human insulin
- compound
- swine
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Expired
Links
Classifications
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
- C07K14/435—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
- C07K14/575—Hormones
- C07K14/62—Insulins
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P3/00—Drugs for disorders of the metabolism
- A61P3/08—Drugs for disorders of the metabolism for glucose homeostasis
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P3/00—Drugs for disorders of the metabolism
- A61P3/08—Drugs for disorders of the metabolism for glucose homeostasis
- A61P3/10—Drugs for disorders of the metabolism for glucose homeostasis for hyperglycaemia, e.g. antidiabetics
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K38/00—Medicinal preparations containing peptides
Abstract
Abstract of the Disclosure The invention relates to an insulin of the formula I
Description
17~3 The invention relates -to insu:l ins of the formula I
.
Gly (A1 Ile Yal ( B2 ) Yal Asn ~lu Gln Gln Hi.s , ~--Cys - IJeu , Cys--S_ S_ Cys 10 Thr Giy Se r . Se r Ile His : i--Cys Leu Ser Yal 15I,eu Glu Tyr Ala G;n Leu I.eu Tyr -Glu Leu ~0.Asn ~al .
Tyr / S Cys .
- Cys - - S Gly Y ~A2 1 ) . Glu Arg . C;ly Phe Phe Tyr Thr 3 Pro Lys ~B29) ' ~.
~ :~ 7 3 ~3 ~3 ~ part from ~ small modification, the new compounds have the am~noacid sequence of human insulin Insulins in which the Bl-phenylalanine is missing are already known, but desBl-phenylalanine-human insulin (des-PheBl-human insulin) has not hitherto been described.
Since human insulin can now be prepared in several ways as a starting material for experimental purposes, it has for the first time been possible to recognize the advantageous properties ofdes-PheBl-human insulin (formula I, in which X = Thr and Y = Asn).
The solubility of the compounds accor~ing to the invention in which X = Thr is/ quite generally, con$ider-ably greater than that of compounds which still contain PheBl, so that highly concentrated solutions such as are 15 required for insulin pumps can be prepared Moreover, these compounds have advantages in the immunological field inasmuch as they glve rise to a lesser degree of formation of antibodies. As is known, the production of antibodies, which can even be induced by 20 homologous insulin, plays a critical role in long-term treatment with insulins. Thus, for example, it has been observed that human insulin is al)so capable of pro-ducing antibodies in humans.
A sensitive test model for the production of anti-25 bodies is the goat, which very rapidly forms antibodiesagainst swine insulin and against human insulin in the presence of Freund t S adjuvan-t. According to Diabetes 27 (197~), page 14, figure 4, the bonding capacity of antibodies against the heterologous des-PheBl-swine 1 ~338 4 _ insulin is lower and antibody formation takes place more slowly than in the case of non-modified swine insulin or even in the case of homologous sheep insulin. In this test model, -the compounds according to the invention, above all those in which X = OH, cause an even slower antibody formation, and the bonding capacity is reduced by a further 15 %, compared with that of des-PheBl-swine insulin In a similar manner to human insulin, the compounds 10 according to the invention also exhibit the advantage of a better glucose tolerance, compared with swine insulin, in the treatment of humans. This property is evidently unchanged by spli~ting off PheBl.
- MoreoverJ des-Phe31-human insulin (I in which 15 X = Thr), in particular, has an action which starts very rapidly and is long-lasting.
If ThrB~O ~I in which X - OH~ is also split off enzymatically from the human insulin, in addition -to PheBl, all the advantageous properties mentioned for 20 des-PheBl-human insulin remain. The same applies to the compound which carries, instead of AsnA21, an addi-tional carboxyl group with AspA21 (I in which X = OH and Y = Asp). In this case, an even further increase in the solubility, which manifests i-tself, above all, in 25 the neutral and weakly alkaline range, occurs The particular economic advantage of the compoun~ I in which X _ OH is that human insulin, which at present is still accessible only with grea-t difficulty, does no-t have to be used in the preparation, but -that , .~
swine insulin can be used. Swine insulin differs from human insulin only in the aminoacid B30 (Ala instead oE Thr), and, after this aminoacid has been split off, becomes identical to des-Thr -human insulin.
The splitting off of Ala 30 from swine insulin has already been described in U.S. Patent Specification 3,364,116. However, in respect of its immunological properties, des-Ala 30-swine insulin differs only slightly from swine insulin. In contrast, the additional removal of PheBl causes structural changes in the insulin molecule which are responsible for the advantageous biological properties.
The invention also relates to a process for the pre-paration of the insulins of the formula I. This comprises a) con-verting human insulin, desamidoA21-human insulin, des-ThrB30-human insulin, desamidoA21-des-ThrB30-human insulin, des-Ala 30-swine insulin or desamidoA21-des-AlaB30-swine insulin into the N Al-NEB29-bis-Boc compound and subjecting this to Edman deyradation, b) treating des-PheBl-human insulin or des-PheB30 swine insulin with acid, or c) treating a compound of the formula I in which X denotes Ala with carboxypeptidase A.
To prepare the compounds according to the invention in which X = Thr and Y = Asn, human insulin, which is converted into the corresponding des-PheBl-compound by a procedure analo-gous to that in German Patent 2,005,658, is used as the starting material.
To prepare the compound I in which ;~ =OH and Y = Asn, des-Ala~0-swine insulin, which is prepared, for example, according to Hoppe-Seyler's Z. Physiol.
Chem 359 {1978), page 799t by enzymatic splitting off of AlaB30, is used as the starting ma-terial, and PheBl is split off by a procedure analogous to that in German Patent 2,005,658. It is also possible to reverse the sequence of the steps and first to split off PheBl chemically and then to split off AlaB30 enzymatically.
If Y is to be Asp, the starting compounds are first treated with aqueous acid at pH 2 - 3 and purified by ion exchange chromatography, in a known manner, PheBl or AlaB30 are then re~oved as described above.
It is also possible to carr~ out the acid treat-ment after PheBl and, if appropriate, AlaB30 have been split o~f. This has the advantage that a very uni-form product appears in the final purification by ion exchangerchromatography. 1 - 2 purification operations are thus su ficient, whilst a further purification step is usually required in the reverse process.
The new human insulin analbgues are used for the treatment of diabetes mellitus and, because of their very good solubility at pH 7 - 7.5, can also be used in the insulin pump in a concentration of about 10 %.
The dose depends on the condition of the pa-tient and approximately corresponds to tha-t of swine insulin, but can be considerably lower in the case of patients with an increased level of antibodies.
The invention also relates to a pharmaceutical formulation of the compounds of the formula I which , ~ 1 ~33~3~
contains a compound of the formula I in dissolved or amorphous form, in addi-tion to a compound of the formula I, in crystalline form, which howeve~, addltionally con-tains phenylalanine ln Bl.
The new human insulin analogues can, of course, also be used together with compounds which still contain PheBl. They are then in the dissolved or amorphous form, whilst the compounds containing PheBl are employed in crystalline form. Two-phase insulins in which a rapid onset of action is combined with a long period of action are known per se. They are des-cribed, for.example, in German Offenle~gungsschrift
.
Gly (A1 Ile Yal ( B2 ) Yal Asn ~lu Gln Gln Hi.s , ~--Cys - IJeu , Cys--S_ S_ Cys 10 Thr Giy Se r . Se r Ile His : i--Cys Leu Ser Yal 15I,eu Glu Tyr Ala G;n Leu I.eu Tyr -Glu Leu ~0.Asn ~al .
Tyr / S Cys .
- Cys - - S Gly Y ~A2 1 ) . Glu Arg . C;ly Phe Phe Tyr Thr 3 Pro Lys ~B29) ' ~.
~ :~ 7 3 ~3 ~3 ~ part from ~ small modification, the new compounds have the am~noacid sequence of human insulin Insulins in which the Bl-phenylalanine is missing are already known, but desBl-phenylalanine-human insulin (des-PheBl-human insulin) has not hitherto been described.
Since human insulin can now be prepared in several ways as a starting material for experimental purposes, it has for the first time been possible to recognize the advantageous properties ofdes-PheBl-human insulin (formula I, in which X = Thr and Y = Asn).
The solubility of the compounds accor~ing to the invention in which X = Thr is/ quite generally, con$ider-ably greater than that of compounds which still contain PheBl, so that highly concentrated solutions such as are 15 required for insulin pumps can be prepared Moreover, these compounds have advantages in the immunological field inasmuch as they glve rise to a lesser degree of formation of antibodies. As is known, the production of antibodies, which can even be induced by 20 homologous insulin, plays a critical role in long-term treatment with insulins. Thus, for example, it has been observed that human insulin is al)so capable of pro-ducing antibodies in humans.
A sensitive test model for the production of anti-25 bodies is the goat, which very rapidly forms antibodiesagainst swine insulin and against human insulin in the presence of Freund t S adjuvan-t. According to Diabetes 27 (197~), page 14, figure 4, the bonding capacity of antibodies against the heterologous des-PheBl-swine 1 ~338 4 _ insulin is lower and antibody formation takes place more slowly than in the case of non-modified swine insulin or even in the case of homologous sheep insulin. In this test model, -the compounds according to the invention, above all those in which X = OH, cause an even slower antibody formation, and the bonding capacity is reduced by a further 15 %, compared with that of des-PheBl-swine insulin In a similar manner to human insulin, the compounds 10 according to the invention also exhibit the advantage of a better glucose tolerance, compared with swine insulin, in the treatment of humans. This property is evidently unchanged by spli~ting off PheBl.
- MoreoverJ des-Phe31-human insulin (I in which 15 X = Thr), in particular, has an action which starts very rapidly and is long-lasting.
If ThrB~O ~I in which X - OH~ is also split off enzymatically from the human insulin, in addition -to PheBl, all the advantageous properties mentioned for 20 des-PheBl-human insulin remain. The same applies to the compound which carries, instead of AsnA21, an addi-tional carboxyl group with AspA21 (I in which X = OH and Y = Asp). In this case, an even further increase in the solubility, which manifests i-tself, above all, in 25 the neutral and weakly alkaline range, occurs The particular economic advantage of the compoun~ I in which X _ OH is that human insulin, which at present is still accessible only with grea-t difficulty, does no-t have to be used in the preparation, but -that , .~
swine insulin can be used. Swine insulin differs from human insulin only in the aminoacid B30 (Ala instead oE Thr), and, after this aminoacid has been split off, becomes identical to des-Thr -human insulin.
The splitting off of Ala 30 from swine insulin has already been described in U.S. Patent Specification 3,364,116. However, in respect of its immunological properties, des-Ala 30-swine insulin differs only slightly from swine insulin. In contrast, the additional removal of PheBl causes structural changes in the insulin molecule which are responsible for the advantageous biological properties.
The invention also relates to a process for the pre-paration of the insulins of the formula I. This comprises a) con-verting human insulin, desamidoA21-human insulin, des-ThrB30-human insulin, desamidoA21-des-ThrB30-human insulin, des-Ala 30-swine insulin or desamidoA21-des-AlaB30-swine insulin into the N Al-NEB29-bis-Boc compound and subjecting this to Edman deyradation, b) treating des-PheBl-human insulin or des-PheB30 swine insulin with acid, or c) treating a compound of the formula I in which X denotes Ala with carboxypeptidase A.
To prepare the compounds according to the invention in which X = Thr and Y = Asn, human insulin, which is converted into the corresponding des-PheBl-compound by a procedure analo-gous to that in German Patent 2,005,658, is used as the starting material.
To prepare the compound I in which ;~ =OH and Y = Asn, des-Ala~0-swine insulin, which is prepared, for example, according to Hoppe-Seyler's Z. Physiol.
Chem 359 {1978), page 799t by enzymatic splitting off of AlaB30, is used as the starting ma-terial, and PheBl is split off by a procedure analogous to that in German Patent 2,005,658. It is also possible to reverse the sequence of the steps and first to split off PheBl chemically and then to split off AlaB30 enzymatically.
If Y is to be Asp, the starting compounds are first treated with aqueous acid at pH 2 - 3 and purified by ion exchange chromatography, in a known manner, PheBl or AlaB30 are then re~oved as described above.
It is also possible to carr~ out the acid treat-ment after PheBl and, if appropriate, AlaB30 have been split o~f. This has the advantage that a very uni-form product appears in the final purification by ion exchangerchromatography. 1 - 2 purification operations are thus su ficient, whilst a further purification step is usually required in the reverse process.
The new human insulin analbgues are used for the treatment of diabetes mellitus and, because of their very good solubility at pH 7 - 7.5, can also be used in the insulin pump in a concentration of about 10 %.
The dose depends on the condition of the pa-tient and approximately corresponds to tha-t of swine insulin, but can be considerably lower in the case of patients with an increased level of antibodies.
The invention also relates to a pharmaceutical formulation of the compounds of the formula I which , ~ 1 ~33~3~
contains a compound of the formula I in dissolved or amorphous form, in addi-tion to a compound of the formula I, in crystalline form, which howeve~, addltionally con-tains phenylalanine ln Bl.
The new human insulin analogues can, of course, also be used together with compounds which still contain PheBl. They are then in the dissolved or amorphous form, whilst the compounds containing PheBl are employed in crystalline form. Two-phase insulins in which a rapid onset of action is combined with a long period of action are known per se. They are des-cribed, for.example, in German Offenle~gungsschrift
2,4~9,515 (HOE 74/F 377) and German Offenlegungsschrift 2,256,215 ~HOE 72/F 348). The new ~ormulations accor-ding to the invention are prepared analogously to the~nstructions given for swine insulin in these specifica-tions. The invention also relates to another formula~
tion form in which a compound o~ the ~ormula I in which Y = Asp is present in solution in addition to the corres-po~ding compound in which Y = Asn, in crystalline form.
This formulation form can be prepared by a procedure analogGus to that in German Offenlegungsschrift 2,803,9~6.
EXAMPLES
The intermediate and end produc-ts were characteri-zed by paper electrophoresis and gel electrophoresis a-t pH 2 and 8.3, and by aminoacid analysis, thin layer chroma-tography and HPLC, in the known systems.
Example 1:
~ ~ 73388 _ ~ _ Des-PheBl-h~lman insulin a) 1 g of human insulin is taken up in 70 ml of 80 % streng-th aqueous dimethylacetarnide. 1.45 g of tert.-butoxycarbonyl azide and 3.3 ml of lN sodium bi-carbonate are added and the mixture is stirred at 3~ Cfor 5 hours. The solution is then concentrated at a bath temperature of at most 50C. ,The residue is triturated with ether and digested with 10 rnl of 2 /0 strength acetic acid. Yield: 942 mg of NaAl, N~B29-di-tert.-butoxycarbonyl-hùman insulin.
b) The resulting compound is dissolved in 4 cc of 95 % strength pyridine, 0.03 cc of phenyl isothio-cyanate are added and mixture is stirred at room tempera-ture for 4 hours~ It is then concentrated to a small volume in vacuo at a bath temperature of at mast 50C, and the compound is precipitated with ether. Yield:
'830 of NaAl ~ B29-di-tert,-butoxycarbonyl-M -phenylthiocarbamoyl-human insulin.
c) 820 mg of the compound prepared according to b) are kep-t in 8.5 cc of trifluoroacetic acid at room temperature for 1 hour, When 100 cc of ether are added, 725 mg of des-phenylalanineBl-human insulin pre-cipita-te; the product can be made to crystallize at pH 5.0 -to 5,5 in the known manner, Phe calculated 2.00, found 2.02 Example 2:
Des-PheBl-des-ThrB30-human insu].in a) 1,0 g of des-AlaB30-swine insulin, prepared accor-' ding to Hoppe-Seyler's Z. Physiol. Chem. 359 (1978), 733~3~
_ g _ page 799, are reacted by a procedure analogous to that in Example la c. 710 mg of the title compound are obtained and can be purified by chromatography in 1 %
strength acetic acid on a Sephadex G 50 superfine 1 x 5 100 cm column. Yield: 623 mg. Crystallization at pH 5.4.
Phe calcula-ted 2.00, *ound 1.99 b) 50b mg of des-PheBl-swine insulin, prepared from swine insulin by a procedure analogous to tha-t in 10 Example 1, are dissolved in 100 ml of 0,lM ammonium bicarbonate buffer at pH 8.2. 3 mg of carboxypeptid-ase A are added and the mixture is kept at room tempera-ture for 15 hours and then lyophilized. To remove salts, the product is chromatographed in 1 % streng-th acetic acid over a Sephadex G 15 1 x 50 cm column.
For further purification, the product was chroma-tographed in 30 % strength isopropanol containing 0.05M
of tris-buffer a-t pH 8 and with a NaCl gradient of 0 to 0.25M in a DEAE-Sephadex A-25 2.5 x 5~ cm column.
The eluate was deszlinated by dialysis against distilled water and lyophilized. Yield: 276 mg, Phe calcula-ted 2, found 1.98; Ala calculated 1, found 1.01 _ample ~:
Des-Phe31-rAspA21]-human insulin a~ 0.5 g of [AspA21]-human insulin, prepared by keeping human insulin in aqueous trifluoroacetic acid at pH 2 for 3 days and freeze-drying the product and purifying it by chromatography on DEAE-Sephadex A-25 ~d~n~1les ~r~ rk .~ .
7~33~
by a procedure analogous to that in Example 2b), is reacted by a procedure analogous to that in Example la -c).
A~ter chromatography by a procedure analogous to that in Example 2a), 324 mg of the title compound are obtained.
Phe calculated 2, found 2.02 The product is characterized by gel electrophoresis - at pH 8. The distance -travelled corresponds to that in the case of desamido-insulin (slight deviation as a - result of the absence of phenylalanine).
b) 005 g f des-PheBl-human insulin, prepared accor-ding to Example 1, are kept in aqueous trifluoroacetic acid as described under a) and then purified on DEAE-Sephadex A-25. Crystalliza-tion is carried out in the known manner at pH 5.0 to gi.ve 365 mg of the ti-tle com-1~ pound, which is identical to that prepared according toa) Exam~le 4-t Des-PheBl-des-ThrB30-LAspA21]-human insulin 0.5 g of des-PheBl-des-Thr330-human insulin, prepared according to Example 2, is treated with acid and purified, according to Example 3b). Yield:
344 mg Phe calcula-ted 2, found 1.99; Ala calculated 1, ~ound 1.02 The product is fur-ther characterized by gel electro-phoresis at pH 8, by a procedure analogous to that in Example 3a).
tion form in which a compound o~ the ~ormula I in which Y = Asp is present in solution in addition to the corres-po~ding compound in which Y = Asn, in crystalline form.
This formulation form can be prepared by a procedure analogGus to that in German Offenlegungsschrift 2,803,9~6.
EXAMPLES
The intermediate and end produc-ts were characteri-zed by paper electrophoresis and gel electrophoresis a-t pH 2 and 8.3, and by aminoacid analysis, thin layer chroma-tography and HPLC, in the known systems.
Example 1:
~ ~ 73388 _ ~ _ Des-PheBl-h~lman insulin a) 1 g of human insulin is taken up in 70 ml of 80 % streng-th aqueous dimethylacetarnide. 1.45 g of tert.-butoxycarbonyl azide and 3.3 ml of lN sodium bi-carbonate are added and the mixture is stirred at 3~ Cfor 5 hours. The solution is then concentrated at a bath temperature of at most 50C. ,The residue is triturated with ether and digested with 10 rnl of 2 /0 strength acetic acid. Yield: 942 mg of NaAl, N~B29-di-tert.-butoxycarbonyl-hùman insulin.
b) The resulting compound is dissolved in 4 cc of 95 % strength pyridine, 0.03 cc of phenyl isothio-cyanate are added and mixture is stirred at room tempera-ture for 4 hours~ It is then concentrated to a small volume in vacuo at a bath temperature of at mast 50C, and the compound is precipitated with ether. Yield:
'830 of NaAl ~ B29-di-tert,-butoxycarbonyl-M -phenylthiocarbamoyl-human insulin.
c) 820 mg of the compound prepared according to b) are kep-t in 8.5 cc of trifluoroacetic acid at room temperature for 1 hour, When 100 cc of ether are added, 725 mg of des-phenylalanineBl-human insulin pre-cipita-te; the product can be made to crystallize at pH 5.0 -to 5,5 in the known manner, Phe calculated 2.00, found 2.02 Example 2:
Des-PheBl-des-ThrB30-human insu].in a) 1,0 g of des-AlaB30-swine insulin, prepared accor-' ding to Hoppe-Seyler's Z. Physiol. Chem. 359 (1978), 733~3~
_ g _ page 799, are reacted by a procedure analogous to that in Example la c. 710 mg of the title compound are obtained and can be purified by chromatography in 1 %
strength acetic acid on a Sephadex G 50 superfine 1 x 5 100 cm column. Yield: 623 mg. Crystallization at pH 5.4.
Phe calcula-ted 2.00, *ound 1.99 b) 50b mg of des-PheBl-swine insulin, prepared from swine insulin by a procedure analogous to tha-t in 10 Example 1, are dissolved in 100 ml of 0,lM ammonium bicarbonate buffer at pH 8.2. 3 mg of carboxypeptid-ase A are added and the mixture is kept at room tempera-ture for 15 hours and then lyophilized. To remove salts, the product is chromatographed in 1 % streng-th acetic acid over a Sephadex G 15 1 x 50 cm column.
For further purification, the product was chroma-tographed in 30 % strength isopropanol containing 0.05M
of tris-buffer a-t pH 8 and with a NaCl gradient of 0 to 0.25M in a DEAE-Sephadex A-25 2.5 x 5~ cm column.
The eluate was deszlinated by dialysis against distilled water and lyophilized. Yield: 276 mg, Phe calcula-ted 2, found 1.98; Ala calculated 1, found 1.01 _ample ~:
Des-Phe31-rAspA21]-human insulin a~ 0.5 g of [AspA21]-human insulin, prepared by keeping human insulin in aqueous trifluoroacetic acid at pH 2 for 3 days and freeze-drying the product and purifying it by chromatography on DEAE-Sephadex A-25 ~d~n~1les ~r~ rk .~ .
7~33~
by a procedure analogous to that in Example 2b), is reacted by a procedure analogous to that in Example la -c).
A~ter chromatography by a procedure analogous to that in Example 2a), 324 mg of the title compound are obtained.
Phe calculated 2, found 2.02 The product is characterized by gel electrophoresis - at pH 8. The distance -travelled corresponds to that in the case of desamido-insulin (slight deviation as a - result of the absence of phenylalanine).
b) 005 g f des-PheBl-human insulin, prepared accor-ding to Example 1, are kept in aqueous trifluoroacetic acid as described under a) and then purified on DEAE-Sephadex A-25. Crystalliza-tion is carried out in the known manner at pH 5.0 to gi.ve 365 mg of the ti-tle com-1~ pound, which is identical to that prepared according toa) Exam~le 4-t Des-PheBl-des-ThrB30-LAspA21]-human insulin 0.5 g of des-PheBl-des-Thr330-human insulin, prepared according to Example 2, is treated with acid and purified, according to Example 3b). Yield:
344 mg Phe calcula-ted 2, found 1.99; Ala calculated 1, ~ound 1.02 The product is fur-ther characterized by gel electro-phoresis at pH 8, by a procedure analogous to that in Example 3a).
Claims (6)
OR PRIVILEGE IS CLAIMED ARE DEFINED AS FOLLOWS:
1. A process for the preparation of an insulin of the formula I
(I) wherein X denotes Thr or OH and Y denotes Asp or Asn, in which (a) human insulin, desamidoA21-human insulin, des-ThrB30-human insulin, desamidoA21-des-ThrB30-human insulin, des-AlaB30-swine insulin or desamidoA21-des-AlaB30-swine insulin is converted into the NA1-NEB29-bis-Boc compound and this compound is subjected to Edman degradation, or (b) des-PheB1-human insulin or des-PheB30 swine insulin is treated with acid to produce a compound wherein X = Asp or, (c) a compound of the formula I in which X denotes Ala is treated with carboxypeptidase A.
(I) wherein X denotes Thr or OH and Y denotes Asp or Asn, in which (a) human insulin, desamidoA21-human insulin, des-ThrB30-human insulin, desamidoA21-des-ThrB30-human insulin, des-AlaB30-swine insulin or desamidoA21-des-AlaB30-swine insulin is converted into the NA1-NEB29-bis-Boc compound and this compound is subjected to Edman degradation, or (b) des-PheB1-human insulin or des-PheB30 swine insulin is treated with acid to produce a compound wherein X = Asp or, (c) a compound of the formula I in which X denotes Ala is treated with carboxypeptidase A.
2. A process as claimed in claim 1 in which the preparation is carried out according to reaction (a).
3. A process as claimed in claim 1 in which the preparation is carried out according to reaction (b).
4. A compound of the formula I as defined in claim 1, whenever obtained according to a process as claimed in claim 1, claim 2 or claim 3 or by an obvious chemical equivalent thereof.
5. A process as claimed in claim 1 in which the preparation is carried out according to reaction (c).
6. A compound of the formula I as defined in claim 1, whenever obtained according to a process as claimed in claim 5 or by an obvious chemical equivalent thereof.
Applications Claiming Priority (2)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| DE19803033127 DE3033127A1 (en) | 1980-09-03 | 1980-09-03 | NEW INSULIN ANALOG |
| DEP3033127.2 | 1980-09-03 |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CA1173388A true CA1173388A (en) | 1984-08-28 |
Family
ID=6111025
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CA000385067A Expired CA1173388A (en) | 1980-09-03 | 1981-09-02 | Analogues of insulin |
Country Status (9)
| Country | Link |
|---|---|
| EP (1) | EP0046979B1 (en) |
| JP (1) | JPS5777655A (en) |
| AT (1) | ATE4591T1 (en) |
| AU (1) | AU545399B2 (en) |
| CA (1) | CA1173388A (en) |
| DE (2) | DE3033127A1 (en) |
| DK (1) | DK149778C (en) |
| ES (1) | ES8206448A1 (en) |
| ZA (1) | ZA816085B (en) |
Cited By (12)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US5015728A (en) * | 1983-09-17 | 1991-05-14 | Hoechst Aktiengesellschaft | Process for the preparation of insulin derivatives, the B chain of which is lengthened c-terminally |
| US7476652B2 (en) | 2002-06-18 | 2009-01-13 | Sanofi-Aventis Deutschland Gmbh | Acidic insulin preparations having improved stability |
| US9364519B2 (en) | 2011-09-01 | 2016-06-14 | Sanofi-Aventis Deutschland Gmbh | Pharmaceutical composition for use in the treatment of a neurodegenerative disease |
| US9408893B2 (en) | 2011-08-29 | 2016-08-09 | Sanofi-Aventis Deutschland Gmbh | Pharmaceutical combination for use in glycemic control in diabetes type 2 patients |
| US9526764B2 (en) | 2008-10-17 | 2016-12-27 | Sanofi-Aventis Deutschland Gmbh | Combination of an insulin and a GLP-1-agonist |
| US9707176B2 (en) | 2009-11-13 | 2017-07-18 | Sanofi-Aventis Deutschland Gmbh | Pharmaceutical composition comprising a GLP-1 agonist and methionine |
| US9821032B2 (en) | 2011-05-13 | 2017-11-21 | Sanofi-Aventis Deutschland Gmbh | Pharmaceutical combination for improving glycemic control as add-on therapy to basal insulin |
| US9950039B2 (en) | 2014-12-12 | 2018-04-24 | Sanofi-Aventis Deutschland Gmbh | Insulin glargine/lixisenatide fixed ratio formulation |
| US9981013B2 (en) | 2010-08-30 | 2018-05-29 | Sanofi-Aventis Deutschland Gmbh | Use of AVE0010 for the treatment of diabetes mellitus type 2 |
| US10029011B2 (en) | 2009-11-13 | 2018-07-24 | Sanofi-Aventis Deutschland Gmbh | Pharmaceutical composition comprising a GLP-1 agonist, an insulin and methionine |
| US10159713B2 (en) | 2015-03-18 | 2018-12-25 | Sanofi-Aventis Deutschland Gmbh | Treatment of type 2 diabetes mellitus patients |
| US10434147B2 (en) | 2015-03-13 | 2019-10-08 | Sanofi-Aventis Deutschland Gmbh | Treatment type 2 diabetes mellitus patients |
Families Citing this family (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| DE3327928A1 (en) * | 1983-08-03 | 1985-02-21 | Hoechst Ag, 6230 Frankfurt | METHOD FOR PRODUCING INSULIN DERIVATIVES |
| DK347086D0 (en) * | 1986-07-21 | 1986-07-21 | Novo Industri As | NOVEL PEPTIDES |
| DE3837825A1 (en) * | 1988-11-08 | 1990-05-10 | Hoechst Ag | NEW INSULIN DERIVATIVES, THEIR USE AND A PHARMACEUTICAL PREPARATION CONTAINING THEM |
| CN1125081C (en) | 1999-09-08 | 2003-10-22 | 中国科学院上海生物化学研究所 | Recombined natural and new-type human insulin and its preparation |
| DE10114178A1 (en) | 2001-03-23 | 2002-10-10 | Aventis Pharma Gmbh | Zinc-free and low-zinc insulin preparations with improved stability |
Family Cites Families (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US3364116A (en) * | 1964-11-02 | 1968-01-16 | Squibb & Sons Inc | Method of use of desalanino insulin compositions |
| DE2256215C3 (en) * | 1972-11-16 | 1981-01-08 | Hoechst Ag, 6000 Frankfurt | Insulin preparation containing crystalline insulin and amorphous desphenylalanine8 '-insulin |
-
1980
- 1980-09-03 DE DE19803033127 patent/DE3033127A1/en not_active Withdrawn
-
1981
- 1981-08-26 EP EP81106625A patent/EP0046979B1/en not_active Expired
- 1981-08-26 AT AT81106625T patent/ATE4591T1/en not_active IP Right Cessation
- 1981-08-26 DE DE8181106625T patent/DE3160852D1/en not_active Expired
- 1981-08-28 ES ES505039A patent/ES8206448A1/en not_active Expired
- 1981-09-01 JP JP56136212A patent/JPS5777655A/en active Pending
- 1981-09-02 DK DK388081A patent/DK149778C/en active
- 1981-09-02 AU AU74883/81A patent/AU545399B2/en not_active Ceased
- 1981-09-02 ZA ZA816085A patent/ZA816085B/en unknown
- 1981-09-02 CA CA000385067A patent/CA1173388A/en not_active Expired
Cited By (16)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US5015728A (en) * | 1983-09-17 | 1991-05-14 | Hoechst Aktiengesellschaft | Process for the preparation of insulin derivatives, the B chain of which is lengthened c-terminally |
| US7476652B2 (en) | 2002-06-18 | 2009-01-13 | Sanofi-Aventis Deutschland Gmbh | Acidic insulin preparations having improved stability |
| US7713930B2 (en) | 2002-06-18 | 2010-05-11 | Sanofi-Aventis Deutschland Gmbh | Acidic insulin preparations having improved stability |
| US10117909B2 (en) | 2008-10-17 | 2018-11-06 | Sanofi-Aventis Deutschland Gmbh | Combination of an insulin and a GLP-1 agonist |
| US9526764B2 (en) | 2008-10-17 | 2016-12-27 | Sanofi-Aventis Deutschland Gmbh | Combination of an insulin and a GLP-1-agonist |
| US10029011B2 (en) | 2009-11-13 | 2018-07-24 | Sanofi-Aventis Deutschland Gmbh | Pharmaceutical composition comprising a GLP-1 agonist, an insulin and methionine |
| US9707176B2 (en) | 2009-11-13 | 2017-07-18 | Sanofi-Aventis Deutschland Gmbh | Pharmaceutical composition comprising a GLP-1 agonist and methionine |
| US10028910B2 (en) | 2009-11-13 | 2018-07-24 | Sanofi-Aventis Deutschland Gmbh | Pharmaceutical composition comprising a GLP-1-agonist and methionine |
| US9981013B2 (en) | 2010-08-30 | 2018-05-29 | Sanofi-Aventis Deutschland Gmbh | Use of AVE0010 for the treatment of diabetes mellitus type 2 |
| US9821032B2 (en) | 2011-05-13 | 2017-11-21 | Sanofi-Aventis Deutschland Gmbh | Pharmaceutical combination for improving glycemic control as add-on therapy to basal insulin |
| US9408893B2 (en) | 2011-08-29 | 2016-08-09 | Sanofi-Aventis Deutschland Gmbh | Pharmaceutical combination for use in glycemic control in diabetes type 2 patients |
| US9987332B2 (en) | 2011-09-01 | 2018-06-05 | Sanofi-Aventis Deutschland Gmbh | Pharmaceutical composition for use in the treatment of a neurodegenerative disease |
| US9364519B2 (en) | 2011-09-01 | 2016-06-14 | Sanofi-Aventis Deutschland Gmbh | Pharmaceutical composition for use in the treatment of a neurodegenerative disease |
| US9950039B2 (en) | 2014-12-12 | 2018-04-24 | Sanofi-Aventis Deutschland Gmbh | Insulin glargine/lixisenatide fixed ratio formulation |
| US10434147B2 (en) | 2015-03-13 | 2019-10-08 | Sanofi-Aventis Deutschland Gmbh | Treatment type 2 diabetes mellitus patients |
| US10159713B2 (en) | 2015-03-18 | 2018-12-25 | Sanofi-Aventis Deutschland Gmbh | Treatment of type 2 diabetes mellitus patients |
Also Published As
| Publication number | Publication date |
|---|---|
| AU7488381A (en) | 1982-03-11 |
| DK149778B (en) | 1986-09-29 |
| ZA816085B (en) | 1982-08-25 |
| EP0046979A1 (en) | 1982-03-10 |
| ES505039A0 (en) | 1982-08-16 |
| ES8206448A1 (en) | 1982-08-16 |
| DK388081A (en) | 1982-03-04 |
| DE3033127A1 (en) | 1982-04-08 |
| ATE4591T1 (en) | 1983-09-15 |
| JPS5777655A (en) | 1982-05-15 |
| EP0046979B1 (en) | 1983-09-07 |
| DK149778C (en) | 1987-04-06 |
| DE3160852D1 (en) | 1983-10-13 |
| AU545399B2 (en) | 1985-07-11 |
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