NZ232808A - Specifically glycosylated insulin derivatives with between one and two sugar units - Google Patents
Specifically glycosylated insulin derivatives with between one and two sugar unitsInfo
- Publication number
- NZ232808A NZ232808A NZ232808A NZ23280890A NZ232808A NZ 232808 A NZ232808 A NZ 232808A NZ 232808 A NZ232808 A NZ 232808A NZ 23280890 A NZ23280890 A NZ 23280890A NZ 232808 A NZ232808 A NZ 232808A
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- NZ
- New Zealand
- Prior art keywords
- insulin
- phe
- human insulin
- glycosylated
- human
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-
- 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
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K38/00—Medicinal preparations containing peptides
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- Life Sciences & Earth Sciences (AREA)
- Biochemistry (AREA)
- Genetics & Genomics (AREA)
- Gastroenterology & Hepatology (AREA)
- Toxicology (AREA)
- Endocrinology (AREA)
- Biophysics (AREA)
- General Health & Medical Sciences (AREA)
- Zoology (AREA)
- Medicinal Chemistry (AREA)
- Molecular Biology (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Diabetes (AREA)
- Medicines That Contain Protein Lipid Enzymes And Other Medicines (AREA)
- Peptides Or Proteins (AREA)
- Medicinal Preparation (AREA)
Description
<div id="description" class="application article clearfix">
<p lang="en" class="printTableText">New Zealand Paient Spedficaiion for Paient Number £32808 <br><br>
232808 <br><br>
Priority Date(s): ... .w?.. <br><br>
ilod: ...iv?..T.SjO.... <br><br>
I <br><br>
| '.icMcatiSfl Dvte: 2.8.AP.R. <br><br>
| P.O. Journal-No: <br><br>
sm, £ <br><br>
Oomr»^?fi Specification F <br><br>
™ .t K- ^ ff <br><br>
^ » £ If'l? <br><br>
\ <br><br>
Patents Form No. 5 <br><br>
NEW ZEALAND PATENTS ACT 1953 COMPLETE SPECIFICATION <br><br>
PEPTIDES <br><br>
WE, NOVO NORDISK A/S, a Danish joint-stock company of Novo Alle, 2 880 Bagsvaerd, Denmark hereby declare the invention, for which we pray that a patent may be granted to us, and the method by which it is to be performed, to be particularly described in and by the following statement: <br><br>
- 1 - <br><br>
(followed by page la) <br><br>
ATS/ToN/VMN/VNy/KGF, 1990-02-23, IL <br><br>
3290.2 <br><br>
23 2 8 08 <br><br>
The present invention relates to specifically glycosylated insulins and combinations thereof, pharmaceutical preparations containing such glycosylated compounds and a method for their preparation. <br><br>
In the recent years, several insulin analogues have been suggested for the treatment of diabetes mellitus. The purpose of developing such insulin analogues has been to improve the insulin replacement therapy by making available insulin analogues with either a more rapid or a more protracted insulin action compared to especially human insulin. <br><br>
A problem in the development of insulin analogues by substituting one or more of the amino acid residues in native insulin is the potential immunogenicity of such compounds. Also unforeseeable solubility and stability problems may arise from such substitutions. <br><br>
Although insulin has a very short half life time in circulating blood, it can not be excluded that a small amount of insulin is glycosylated in vivo not only in diabetic patients as postulated by Nakayama et al. (Nonenzymatic glyco-sylation of insulin in "Current topics in clinical and experimental aspects of diabetes mellitus" (1985), 201 - 204, Sakamoto, Min and Baba, Eds., Elsevier Science Publishers B.V.) but also in non-diabetics. It is therefore possible that the organism has developed mechanisms to suppress the formation of antibodies against glycosylated insulin. It is furthermore possible that the conformational changes of the saccharide part will be able to camouflage the antigen. <br><br>
The binding of glucose, mannose and certain oligosaccharides to insulin has been the subject of numerous in vitro studies in the past with the purpose of investigating whether in vivo formation of glycosylated insulin might be responsible for late complications in diabetic patients. Anzen-bacher et al. (Biochimica et Biophysica Acta 386 (1975), 603-607) studied the binding of D-glucose to insulin by equilibrium dialysis. The binding was not found to be very the specific and <br><br>
(followed by page 2) <br><br>
ATS/ToN/VMN/VNy/KGF, 1990-02-23, IL 3290.200 <br><br>
23280a the average number of glucose molecules bound to the insulin molecule was found to be eight. <br><br>
Interaction of insulin with glucose and mannose was studied by Dolhofer et al. (Febs Letters 100 (1979), 133-5 13 6) . The results indicated that both hexoses were covalently incorporated into the insulin molecule upon incubation in vitro at 37°C. Under the chosen reaction conditions, in average 3.6 ± 0.39 glucose and 5.0 ± 0.43 mannose residues were found to be taken up per molecule of insulin. <br><br>
10 A glucose controlled insulin delivery system was sug gested by Brownlee & Cerami (Science 206 (1979), 1190 - 1191, and Diabetes 32, (1983), 499 - 505) by synthesizing glycosylated insulin derivatives which are able to compete with glucose for binding to lectins. In this study maltose and other oligo-15 saccharides were reacted with insulin. <br><br>
Nakayama et al. (supra) investigated non-enzymatic glycosylation of insulin in vitro and in vivo (diabetic patients) and concluded that glucose was incorporated into the insulin molecule in vivo under pathological conditions. By the 20 in vitro studies, 3 molecules of glucose were found to be incorporated per molecule of insulin. <br><br>
In 1988, Lapolla et al. (Diabetes 37 (1988), 787-791) reported a reduced in vivo biological activity of in vitro glycosylated insulin. Insulin was glycosylated in ambient high 25 glucose concentration according to Dolhofer (supra) in aqueous solution at 37°C for 17 hours at a pH value of 7.4. The incorporation of glucose was found to be in average 2 mol glucose residue/mol insulin. <br><br>
Taking into account that native insulin has three 30 free primary amino groups viz. at position B1 (Phe) , Al (Gly) and B29 (Lys), respectively, it is apparent that the above described glycosylated insulin preparations will all be inhomo-geneous mixtures of glycosylated insulin molecules. <br><br>
Nobody has so far taken any steps to fractionate the 35 above mixture into the individual components. <br><br>
ATS/ToN/VMN/VNy/KGF, 1990-02-23, IL 3290.200 <br><br>
3 232 808 <br><br>
Glycosylated insulin derivatives for self-regulating insulin delivery systems have been described by Kim et al. (Journal of Controlled Release l (1984), 57 - 66, and US patent specification Nos. 4,483,792; 4,478,830; 4,478,746 and 5 4,489,063). In these glycosylated insulins, glucose or mannose is coupled with insulin via a spacer group derived from di-carboxylic acids, acid anhydrides or phenyl amines or a combination thereof. <br><br>
European patent application No. 84200328 having pub-10 lication No. 119,650 relates to galactosyl insulins which like the glycosylated insulins described by Kim (supra) contain a spacer group. <br><br>
It is the purpose of this invention to prepare insulin derivatives having improved properties. More specifically, 15 it is the purpose of the present invention to develop non-im-munogenic insulin derivatives. It is furthermore the purpose of the present invention to develop insulin derivatives with a faster onset of insulin action than native insulin and to improve the solubility of less soluble insulins in order to allow 2 0 the use of highly concentrated solutions, for example in insulin pumps. A still further purpose of this invention is to prepare insulin derivatives with an improved stability against fibrillation. <br><br>
This invention provides a specifically glycosylated <br><br>
2 5 insulin that contains one or two monosaccharide groups or one or two oligosaccharide groups with up to three sugar units. Hereinafter the terms "specifically glycosylated insulin" mean insulin having the carbohydrate substituent in a specific position in the insulin molecule. Surprisingly, such <br><br>
3 0 specifically glycosylated insulins offer certain therapeutical advantages as will be apparent from the following description and examples. <br><br>
Preferably the insulin derivative is either monoglycosylated in position Al, B1 or B29; or diglycosylated in 35 position Al and Bl; Al and B29; or B1 and B29. <br><br>
N.Z. PATENT OFFICE ' <br><br>
10 FEB 1992 <br><br>
ATS/ToN/VMN/VNy/KGF,- 1990-02-23, IL ^29 0,. MO <br><br>
j 2 8 u o <br><br>
4 <br><br>
The glycosylated insulins described by Kim et al. and dealt with above are remote from the insulins of the present invention, which utilizes the aldehyde function of the sugar 5 itself to form a covalent bond to insulin, without the use of artificial spacer groups. A further advantage of the present invention over the insulins of Kim et al. is the retention of the natural charge distribution of the insulin molecule. Thus, the amino groups involved which by the glycosylation reaction 10 (see diagram) are converted into secondary amino groups are still capable of being protonated, as in normal insulin. <br><br>
The present insulin derivatives may in each of the three positions contain a monosaccharide or an oligosaccharide with up to three sugar residues. Suitable monosaccharides are 15 glucose, mannose and galactose. Suitable oligosaccharides are maltose, isomaltose, lactose, maltotriose, melibiose and cello-biose. <br><br>
The specifically glycosylated insulin derivatives of this invention may be used as such for the treatment of dia-20 betes mellitus. With the purpose of monitoring the insulin therapy, selected mixtures of the individual specifically glycosylated compounds may however also be used. <br><br>
As used herein, the expression insulins is meant to cover native forms of insulin such as human, bovine and porcine 25 insulin, but also derivatives thereof wherein one or more amino acid residues have been substituted, added or deleted, compared with native insulin, for example as described in European patent applications having publication Nos. 0194864A and 0214826A. <br><br>
3 0 As mentioned above, native insulins have three poten tial glycosylation sites, namely the two N-terminal amino acid residues in the A- and B-chain and the lysine residue in position B29. It is apparent that the number of potential glycosylation sites in insulin analogues of the above described type 35 may be from two (the two N-terminal residues) and upwards de- <br><br>
fN.Z. PATENT Ol'F'C- <br><br>
10 FEB 1992 <br><br>
received I <br><br>
ATS/ToN/VMN/VNy/KGF, 1990-02-23, IL <br><br>
5 <br><br>
3290.200 <br><br>
23 2 8 0ft pending on how many lysine residues are present in the modified insulin molecule, lysine being the only naturally occurring amino acid with a free primary amino group in the side chain. <br><br>
The glycosylation schematically proceeds according to 5 the following diagramme using D-glucose: <br><br>
ch2oh ch2oh <br><br>
10 <br><br>
OH OH <br><br>
15 D-glucose D-glucose pyranose structure chain structure <br><br>
20 <br><br>
The chain structure is the reactive component <br><br>
CH2OH CH2OH <br><br>
.Jr OH -H20 } Os.NH-" insulin" <br><br>
X'OH y^0 + H2N-" insulin" w |C0H } <br><br>
ho i—r +H20 Ho^i—r <br><br>
25 OH OH <br><br>
, -N >- K HO > <br><br>
30 HO>| r CH2-NH-"insulin" <br><br>
HO <br><br>
1 deoxy-D-fructosyl insulin (glucose insulin) "insulin" designates desamino insulin. <br><br>
35 <br><br>
The above reaction will proceed in an analogous manner with other monosaccharides or oligosaccharides having a free aldehyde group. <br><br>
Specific examples of the present glycosylated insu- <br><br>
40 lins are: <br><br>
Phe(Bl) glucose human insulin, <br><br>
Phe(Bl) mannose human insulin, <br><br>
ATS/ToN/VMN/VNy/KGF, 1990-02-23, IL 3290.200 <br><br>
23 2 8 08 <br><br>
Gly(Al) mannose human insulin, <br><br>
Lys(B29) mannose human insulin, <br><br>
Phe(Bl) galactose human insulin, <br><br>
Gly(Al) galactose human insulin, <br><br>
5 Lys(B29) galactose human insulin, <br><br>
Phe(Bl) maltose human insulin, <br><br>
Phe(Bl) lactose human insulin, <br><br>
Gly(Al) glucose human insulin, <br><br>
Gly(Al) maltose human insulin, <br><br>
10 Gly(Al) lactose human insulin, <br><br>
Lys(B2 9) glucose human insulin, <br><br>
Lys(B2 9) maltose human insulin, <br><br>
Lys(B29) lactose human insulin, <br><br>
Gly(Al),Phe(Bl) diglucose human insulin, <br><br>
15 Gly(Al),Lys(B29) diglucose human insulin, <br><br>
Phe(Bl),Lys(B29) diglucose human insulin, <br><br>
Phe(Bl) isomaltose human insulin, <br><br>
Gly(Al) isomaltose human insulin, <br><br>
Lys(B29) isomaltose human insulin, <br><br>
20 Phe(Bl) maltotriose human insulin, <br><br>
Gly(Al) maltotriose human insulin, <br><br>
Lys(B29) maltotriose human insulin, <br><br>
Gly(Al),Phe(Bl) dimaltose human insulin, <br><br>
Gly(Al),Lys(B29) dimaltose human insulin, <br><br>
25 Phe(Bl),Lys(B29) dimaltose human insulin, <br><br>
Gly(Al),Phe(Bl) dilactose human insulin, <br><br>
Gly(Al),Lys(B29) dilactose human insulin, <br><br>
Phe(Bl),Lys(B29) dilactose human insulin, <br><br>
Gly(Al),Phe(Bl) dimaltotriose human insulin, <br><br>
30 Gly(Al),Lys(B29) dimaltotriose human insulin, <br><br>
Phe(Bl),Lys(B29) dimaltotriose human insulin, <br><br>
Phe(Bl) ,Gly(Al) dimannose human insulin, <br><br>
Phe(Bl) ,Lys(B29) dimannose human insulin, <br><br>
Gly(Al),Lys(B29) dimannose human insulin, <br><br>
35 Phe(Bl),Gly(Al) digalactose human insulin, <br><br>
/0O»kx t <br><br>
ATS/ToN/VMN/VNy/KGF, 1990-02-23, IL <br><br>
7 <br><br>
232808 <br><br>
3290.200 <br><br>
15 <br><br>
Phe(Bl),Lys(B29) digalactose human insulin, Gly(Al),Lys(B29) digalactose human insulin, Phe(Bl),Gly(Al) diisomaltose human insulin, Phe(Bl),Lys(B29) diisomaltose human insulin, Gly(Al),Lys(B29) diisomaltose human insulin, <br><br>
10 <br><br>
Phe(Bl) glucose [Asp810] human insulin, <br><br>
Gly(Al),Phe(Bl) diglucose [Asp310] human insulin. <br><br>
Also, specifically glycosylated insulins from other species such as porcine are interesting. <br><br>
The present glycosylated insulins may be prepared by reacting insulin or an insulin analogue with an excess of the 20 selected monosaccharide or oligosaccharide in a suitable organic or aqueous medium. The temperature may vary from 20 to 60 °C. As organic solvents lower carboxylic acids, for example acetic acid and propionic acid, lower aliphatic alcohols, for example methanol, ethanol and 2-propanol, ethylene glycol and 25 propylene glycol may be used. However, phenols may also be used. <br><br>
The duration of the reaction and the composition of the reaction mixture will depend on whether a mono-, di- or triglycosylated end product is desired. The reaction may con-3 0 veniently be followed by reversed phase high pressure liquid chromatography (hereinafter designated RP HPLC) to determine the point of maximum formation of each of the individual glycosylated products. <br><br>
The reaction is stopped by cooling, for example to 35 -20*C, and the reaction mixture is ^.concentrated to dryness h! -^\,N ;'A ;c 8 ,'992 ;? 3 "I. ;ATS/ToN/VMN/VNy/KGF, 1990-02-23, IL ;8 ;3290.200 ;*3280& <br><br>
whereupon the major components of the reaction mixture are isolated and purified by preparative RP HPLC. After desalting, the products are characterized by fast atom bombardment mass spectrometry (hereinafter designated FAB-MS), quantitative amino acid analysis after borohydride reduction and bioassays. <br><br>
The present glycosylated insulins and mixtures thereof may be substituted for the human or porcine insulin in the insulin preparations heretofore known in the art to prepare novel insulin preparations. Such novel insulin preparations will contain the glycosylated insulin or a pharmaceutical^ acceptable salt thereof in an aqueous solution, preferably at a neutral pH value. Preferably, the aqueous medium is made isotonic, for example with sodium chloride, sodium acetate or glycerol. Furthermore, the aqueous medium may contain zinc ions, buffer components such as acetate or phosphate and a preservative such as m-cresol, methylparaben or phenol. The pH value of the preparation may be adjusted to the desired value and the preparation can be sterilized by filtration. <br><br>
The insulin preparation of the present invention can be used similarly to the use of the known insulin preparations. <br><br>
EXPERIMENTAL PART <br><br>
Example 1 <br><br>
PhefBl) glucose human insulin <br><br>
Human insulin (0.1 mmol) was suspended in methanol (30 ml) and glacial acetic acid (5 ml) was added at ambient temperature. The mixture was gently stirred until the insulin had dissolved. Then, a further quantity of methanol (35 ml) was added and after addition of D-glucose (2.2 mmol), the mixture was gently stirred at 40 °C for 8 hours, by which the title compound became the main component. <br><br>
The solution was concentrated almost to dryness on a rotatory evaporator. The residue was dissolved in water and <br><br>
ATS/ToN/VMN/VNy/KGF, 1990-02-23, IL 3290.200 <br><br>
23 2 8 Oft fractionated by preparative RP HPLC. Column: 16 x 250 mm with 7 /xm 100 A C18 particles. Temperature 30°C. Mobile phase. A: 0.04 M phosphoric acid, 0.2 M sodium sulphate, 10% acetonitrile, pH value adjusted to 2.5 with ethanol amine. B: 50% acetonitrile. <br><br>
The fraction corresponding to the central part of the major peak was desalted and lyophilized. The yield was 0.02 mmol. The product was characterized by FAB-MS and quantitative amino acid analysis after borohydride reduction. The amino acid analysis showed the presence of two phenylalanine residues (i.e. one less phenylalanine residue compared to human insulin) proving substitution in Phe(Bl). The molecular weight-was found to be 5970 (calculated: 5970) . <br><br>
Example 2 <br><br>
PheCB1).Glv(All diglucose human insulin <br><br>
The above compound was prepared as described in Example 1 with the exception that the reaction time was 16 hours instead of 8 hours, by which the title compound became the main component. <br><br>
The yield was 0.06 mmol. <br><br>
The amino acid analysis showed the presence of one less Phe and one less Gly residue proving a substitution in position Al and Bl. <br><br>
The molecular weight measured was 6132 (calculated: <br><br>
6132) . <br><br>
The subcutaneous absorption was measured in pigs by injection of 125j labelled Phe(Bl),Gly(Al) diglucose insulin prepared using the iodate method essentially as described (Jor- <br><br>
gensen et al. , Diabetologia 19 (1980), 546 - 554). The absorp- <br><br>
. . . . . 125 <br><br>
tion rate after subcutaneous injection into pigs of I human <br><br>
125 <br><br>
insulin and I Phe(Bl),Gly(Al) diglucose human insulin is shown in Table 1, below. The t75, T50 and T25 values given in Table 1 are the time (in hours) elapsed from the moment of in- <br><br>
ATS/ToN/VMN/VNy/KGF, 1990-02-23, IL <br><br>
10 <br><br>
3290.200 <br><br>
^3280a jection of the sample until the radioactivity measured at the site of injection has decreased to 75%, 50% and 25%, respectively, of the initial value. It appears from Table l that the glycosylated insulin has a significantly faster absorption than 5 human insulin. <br><br>
TABLE 1 <br><br>
T75 T50 T25 <br><br>
10 human insulin 1.12 2.33 3.69 <br><br>
diglucose human insulin 0.65 1.47 2.7 6 <br><br>
The blood glucose lowering effect of human insulin 15 (Actrapid^M) and Gly(Al),Phe(Bl) diglucose human insulin by subcutaneous injection in pigs (mean of 5 animals) in an amount of 0.1 U/kg appears from Table 2, below. Table 2 gives the values for glucose in mmol/1. <br><br>
20 TABLE 2 <br><br>
Time, <br><br>
Human <br><br>
Dialucose human hours insulin insulin <br><br>
-0.33 <br><br>
5.30 <br><br>
5. 24 <br><br>
0 <br><br>
5.36 <br><br>
5.32 <br><br>
0.33 <br><br>
4 .78 <br><br>
4. 76 <br><br>
0.67 <br><br>
4 . 72 <br><br>
4 . 06 <br><br>
1 <br><br>
4 .12 <br><br>
3.34 <br><br>
1.5 <br><br>
3 . 60 <br><br>
2.98 <br><br>
2 <br><br>
3 . 24 <br><br>
2 . 84 <br><br>
2.5 <br><br>
3 .28 <br><br>
2.96 <br><br>
3 <br><br>
3 .18 <br><br>
3 . 14 <br><br>
4 <br><br>
3 . 34 <br><br>
3 . 96 <br><br>
35 <br><br>
/—S <br><br>
ATS/ToN/VMN/VNy/KGF, 1990-02-23, IL 3290.200 <br><br>
11 <br><br>
10 <br><br>
Z3280& <br><br>
Table 2 shows the fast action of diglucose insulin compared with human insulin. <br><br>
The immune responses in rabbits (mean values for 10 animals) of human insulin, bovine insulin and Gly(Al),Phe(Bl) diglucose human insulin appear from Table 3, below, giving values for percentage binding in rabbit serum. (Method: Schlichtkrull et al. (Horm. Metab. Res. Suppl. ser. 5 (1974), 134 - 143)) . <br><br>
TABLE 3 <br><br>
Time. <br><br>
Human <br><br>
Bovine <br><br>
Dialucose <br><br>
davs insulin insulin insulin <br><br>
15 <br><br>
0 <br><br>
1.9 <br><br>
-0.4 <br><br>
1.2 <br><br>
13 <br><br>
2 . 3 <br><br>
0.6 <br><br>
1.3 <br><br>
27 <br><br>
3.0 <br><br>
28.5 <br><br>
1.5 <br><br>
41 <br><br>
3 . 2 <br><br>
29.7 <br><br>
1.4 <br><br>
55 <br><br>
4.3 <br><br>
30.7 <br><br>
1.1 <br><br>
20 <br><br>
69 <br><br>
3.9 <br><br>
32.7 <br><br>
2 . 2 <br><br>
83 <br><br>
2.4 <br><br>
30.1 <br><br>
1.1 <br><br>
97 <br><br>
2 . 0 <br><br>
32.6 <br><br>
1.5 <br><br>
It appears from Table 3 that diglucose human insulin <br><br>
2 5 has a surprisingly low immune response which corresponds to that of human insulin. <br><br>
Example 3 <br><br>
3 0 The following compounds were prepared analogously. <br><br>
The molecular weight measured by FAB-MS or plasma desorption mass spectroscopy, together with the calculated molecular weight, is given for each of the compounds. <br><br></p>
</div>
Claims (17)
1. A specifically glycosylated insulin (as defined in this specification) that contains one or two monosaccharide groups or one or two oligosaccharide groups with up to three sugar units.<br><br>
2. A glycosylated insulin according to claim 1 containing one monosaccharide group or one oligosaccharide group with up to three sugar units .<br><br>
3. A glycosylated insulin according to claim 2 being monoglycosylated in position Al, Bl or B29.<br><br>
4. A glycosylated insulin according to claim 1 being diglycosylated in position Al and Bl; Al and B29; or Bl and B2 9.<br><br>
5. A glycosylated insulin according to any one of the preceding claims containing Asp in position BIO.<br><br>
6. A glycosylated insulin according to any one of the preceding claims in which the parent insulin species is human.<br><br>
7. Phe (Bl) glucose human insulin.<br><br>
8. Phe (Bl), Gly (Al) diglucose human insulin.<br><br>
9. A glycosylated insulin according to claim 1 and substantially as described in this specification with reference to any one of the insulins listed on pages 5 to 7 and any one of the examples.<br><br>
10. A process for the preparation of a specifically glycosylated insulin according to any one of claims 1 to 9 comprising reacting an appropriate insulin in an aqueous or organic medium with a monosaccharide that has a free aldehyde<br><br> N.Z. „<br><br> 10 FEB 1992<br><br> 14<br><br> >32808<br><br> group or an oligosaccharide that has a free aldehyde group and up to three sugar units; and isolating the desired product from the reaction mixture.<br><br>
11. A process as claimed in claim 10 and substantially as described in this specification with reference to any one of the examples.<br><br>
12. A specifically glycosylated insulin whenever prepared by a process according to claim 10 or claim 11.<br><br>
13. A composition containing at least 90% of a specifically glycosylated insulin according to any one of claims 1 to 9 and 12.<br><br>
14. A composition according to claim 13 containing at least 95% by weight of the specifically glycosylated insulin.<br><br>
15. A composition according to claim 13 containing at least 99% by weight of the specifically glycosylated insulin.<br><br>
16. A pharmaceutical preparation containing a specifically glycosylated insulin according to any one of claims 1 to 9 and 12, or a pharmaceutically acceptable salt thereof, optionally together with pharmaceutically acceptable '• adjuvants and additives and preservatives.<br><br>
17. A pharmaceutical preparation as claimed in claim 16 and substantially as described in this specification.<br><br> NOVO NORDISK A/S<br><br> ; a<br><br> " ^ 1 A !<br><br> Byj their attorneys BALDWIN, SON & CAREY<br><br> </p> </div>
Applications Claiming Priority (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
DK111489A DK111489D0 (en) | 1989-03-08 | 1989-03-08 | PEPTIDES |
Publications (1)
Publication Number | Publication Date |
---|---|
NZ232808A true NZ232808A (en) | 1992-04-28 |
Family
ID=8101165
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
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NZ232808A NZ232808A (en) | 1989-03-08 | 1990-03-06 | Specifically glycosylated insulin derivatives with between one and two sugar units |
Country Status (19)
Country | Link |
---|---|
EP (1) | EP0462192A1 (en) |
JP (1) | JPH04504117A (en) |
KR (1) | KR920701249A (en) |
CN (1) | CN1045586A (en) |
AU (1) | AU638701B2 (en) |
CA (1) | CA2049937A1 (en) |
CS (1) | CS114290A3 (en) |
DD (1) | DD296933A5 (en) |
DK (1) | DK111489D0 (en) |
FI (1) | FI914226A0 (en) |
GR (1) | GR1000604B (en) |
HU (1) | HUT59942A (en) |
IL (1) | IL93674A0 (en) |
NO (1) | NO913517L (en) |
NZ (1) | NZ232808A (en) |
PT (1) | PT93366A (en) |
WO (1) | WO1990010645A1 (en) |
YU (1) | YU45490A (en) |
ZA (1) | ZA901737B (en) |
Families Citing this family (23)
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CA2552043A1 (en) | 2004-01-21 | 2005-08-04 | Novo Nordisk A/S | Transglutaminase mediated conjugation of peptides |
US7597884B2 (en) | 2004-08-09 | 2009-10-06 | Alios Biopharma, Inc. | Hyperglycosylated polypeptide variants and methods of use |
US8846103B2 (en) | 2009-01-28 | 2014-09-30 | Smartcells, Inc. | Exogenously triggered controlled release materials and uses thereof |
WO2010088294A1 (en) | 2009-01-28 | 2010-08-05 | Smartcells, Inc. | Conjugate based systems for controlled drug delivery |
CA2750269A1 (en) | 2009-01-28 | 2010-08-05 | Smartcells, Inc. | Crystalline insulin-conjugates |
US8569231B2 (en) | 2009-03-20 | 2013-10-29 | Smartcells, Inc. | Soluble non-depot insulin conjugates and uses thereof |
AU2010226243A1 (en) | 2009-03-20 | 2011-09-22 | Smartcells, Inc. | Terminally-functionalized conjugates and uses thereof |
EP2598170A4 (en) | 2010-07-28 | 2016-07-06 | Smartcells Inc | Drug-ligand conjugates, synthesis thereof, and intermediates thereto |
JP2013535467A (en) | 2010-07-28 | 2013-09-12 | スマートセルズ・インコーポレイテツド | Recombinantly expressed insulin polypeptide and uses thereof |
JP2013541500A (en) | 2010-07-28 | 2013-11-14 | スマートセルズ・インコーポレイテツド | Recombinant lectins, binding site modified lectins and their uses |
US9624287B2 (en) | 2012-07-17 | 2017-04-18 | Case Western Reserve University | O-linked carbohydrate-modified insulin analogues |
EP2877200B1 (en) * | 2012-07-17 | 2019-05-08 | Case Western Reserve University | O-linked carbohydrate-modified insulin analogues |
AU2013346624B2 (en) | 2012-11-13 | 2018-08-09 | Adocia | Quick-acting insulin formulation including a substituted anionic compound |
CA2890048C (en) | 2012-12-03 | 2022-05-03 | Merck Sharp & Dohme Corp. | O-glycosylated carboxy terminal portion (ctp) peptide-based insulin and insulin analogues |
JP6410790B2 (en) | 2013-03-14 | 2018-10-24 | ザ ボード オブ トラスティーズ オブ ザ レランド スタンフォード ジュニア ユニバーシティー | Mitochondrial aldehyde dehydrogenase-2 modulator and method of use thereof |
AU2014329567B2 (en) | 2013-10-04 | 2019-07-25 | Merck Sharp & Dohme Corp. | Glucose-responsive insulin conjugates |
US9795678B2 (en) | 2014-05-14 | 2017-10-24 | Adocia | Fast-acting insulin composition comprising a substituted anionic compound and a polyanionic compound |
FR3020947B1 (en) | 2014-05-14 | 2018-08-31 | Adocia | AQUEOUS COMPOSITION COMPRISING AT LEAST ONE PROTEIN AND A SOLUBILIZING AGENT, ITS PREPARATION AND ITS USES |
FR3043557B1 (en) | 2015-11-16 | 2019-05-31 | Adocia | RAPID ACID COMPOSITION OF INSULIN COMPRISING A SUBSTITUTED CITRATE |
CN105709207A (en) | 2016-01-29 | 2016-06-29 | 徐宝贞 | Medicine for treating gout |
CN105535927A (en) * | 2016-01-29 | 2016-05-04 | 山东中海制药有限公司 | Medicine used for treating influenza, upper respiratory infection and viral pneumonia |
CN105597080A (en) * | 2016-01-29 | 2016-05-25 | 程潜 | Medicine for treating uremia and urine protein |
CN109562185A (en) | 2016-06-02 | 2019-04-02 | 赛诺菲 | The new conjugate of medicament and the part that albumen can be incuded in conjunction with glucose |
Family Cites Families (4)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US3847890A (en) * | 1971-11-01 | 1974-11-12 | A Green | Acidic monosaccharide-substituted proteins |
US4444683A (en) * | 1982-11-17 | 1984-04-24 | University Of Utah | Glycosylated insulin derivatives |
EP0119650A3 (en) * | 1983-03-21 | 1987-09-30 | THE PROCTER & GAMBLE COMPANY | Galactosyl-insulin conjugates useful in treating diabetics |
HU206890B (en) * | 1986-10-13 | 1993-01-28 | Sandoz Ag | Process for producing sugar-modified somatostatin peptide derivatives and pharmaceutical compositions containing them as active components |
-
1989
- 1989-03-08 DK DK111489A patent/DK111489D0/en not_active Application Discontinuation
-
1990
- 1990-03-06 HU HU8727A patent/HUT59942A/en unknown
- 1990-03-06 KR KR1019910701014A patent/KR920701249A/en not_active Application Discontinuation
- 1990-03-06 NZ NZ232808A patent/NZ232808A/en unknown
- 1990-03-06 JP JP2504720A patent/JPH04504117A/en active Pending
- 1990-03-06 AU AU52807/90A patent/AU638701B2/en not_active Expired - Fee Related
- 1990-03-06 EP EP19900904777 patent/EP0462192A1/en not_active Ceased
- 1990-03-06 CA CA002049937A patent/CA2049937A1/en not_active Abandoned
- 1990-03-06 WO PCT/DK1990/000062 patent/WO1990010645A1/en not_active Application Discontinuation
- 1990-03-07 ZA ZA901737A patent/ZA901737B/en unknown
- 1990-03-07 CN CN90101282A patent/CN1045586A/en active Pending
- 1990-03-07 GR GR900100159A patent/GR1000604B/en unknown
- 1990-03-07 DD DD90338488A patent/DD296933A5/en not_active IP Right Cessation
- 1990-03-07 IL IL93674A patent/IL93674A0/en unknown
- 1990-03-07 YU YU00454/90A patent/YU45490A/en unknown
- 1990-03-07 PT PT93366A patent/PT93366A/en not_active Application Discontinuation
- 1990-03-08 CS CS901142A patent/CS114290A3/en unknown
-
1991
- 1991-09-06 NO NO91913517A patent/NO913517L/en unknown
- 1991-09-06 FI FI914226A patent/FI914226A0/en not_active Application Discontinuation
Also Published As
Publication number | Publication date |
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GR1000604B (en) | 1992-08-26 |
CN1045586A (en) | 1990-09-26 |
AU638701B2 (en) | 1993-07-08 |
AU5280790A (en) | 1990-10-09 |
KR920701249A (en) | 1992-08-11 |
WO1990010645A1 (en) | 1990-09-20 |
EP0462192A1 (en) | 1991-12-27 |
JPH04504117A (en) | 1992-07-23 |
GR900100159A (en) | 1990-07-31 |
IL93674A0 (en) | 1990-12-23 |
NO913517L (en) | 1991-11-06 |
YU45490A (en) | 1991-10-31 |
PT93366A (en) | 1990-11-07 |
NO913517D0 (en) | 1991-09-06 |
DK111489D0 (en) | 1989-03-08 |
HUT59942A (en) | 1992-07-28 |
DD296933A5 (en) | 1991-12-19 |
ZA901737B (en) | 1990-11-28 |
CA2049937A1 (en) | 1990-09-09 |
HU902787D0 (en) | 1991-11-28 |
FI914226A0 (en) | 1991-09-06 |
CS114290A3 (en) | 1992-02-19 |
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