DD261096A1 - PROCESS FOR THE INHALATIVE APPLICATION OF BIOLOGICALLY ACTIVE PEPTIDES - Google Patents

Abstract

The invention relates to a method for the inhalative administration of biologically active peptides in human and veterinary medicine. In the inhalation absorption of biologically active peptides - especially insulin - only a fraction of the effect is achieved compared to the injective application form. An increase in absorption can be achieved by co-using surfactants in the preparation for inhalation. As a particularly suitable surfactant, proteinaceous fatty acid condensates were found for inhalative absorption. Hereby, the injective application of the biologically active peptides comparable effects are found. EFK are highly tolerated by the mucous membranes and do not cause irritation or allergies even during long-term use.

Description

Characteristic of the known technical solutions

In the application of biologically active peptides in human and veterinary medicine can usually be dispensed with the injection, since with other forms of application is not to achieve sufficient efficacy.

The solution to this problem may consist in the absorption of biologically active peptides in sufficient quantity through the mucous membranes through the addition of substances acting as absorption mediators. For the rectal, vaginal and nasal application of biologically active substances, a number of absorption mediators are described. These are mostly nonionic, anionic, cationic or ampholytic surfactants.

The object of the surfactants should be to protect the absorption agent from degradation, to substantially increase its absorption through the mucous membranes and thus to ensure a faster onset of action and a higher bioavailability of the active substance. The surfactant used must have extremely low toxicity, be absolutely non-irritating to the mucous membrane, must not undesirably affect the natural bacterial spread of the mucous membranes, not associate with the drug, adversely affect its effects, and must be well tolerated by the galenical excipients ,

Compounds intended to meet these requirements are assigned to both anionic and cationic as well as nonionic and ampholytic surfactants. For example, DE 2641819 proposes bile acids and their alkali metal salts, alkyl and alkylphenol polyglycol ethers and cationic, anionic and ampholytic surfactants, reference being made in particular to the surfactants (alkyl and acyl amino acids) emanating from amino acids for the latter two surfactant groups.

US 4164573 describes for the same purpose polyethylene oxide stearates, lecithin, amino acid derivatives and bile acids, their derivatives and salts.

Of the surfactant groups mentioned, the surfactants deriving from amino acids are by far the least toxic and are considered to be the most compatible with the mucous membranes.

In DE-OS 3018843, however, it is stated that on the absorption of the peptide insulin amino acid derivatives of different chemical structure only a low activity, the salts even exert any effect.

The defined protein fatty acid condensate (EFK) described in WP A61K 2474195 has proved particularly suitable.

The drugs used with the aid of these absorption mediators could be applied with success rectally, vaginally or nasally. These administrations are still fraught with inconvenience.

Object of the invention

The aim of the invention is to find a method for producing biologically active substances that can be absorbed by inhalation (pulmonary) way.

Explanation of the essence of the invention

Surprisingly, it has been found that the protein fatty acid condensates described in WP 2474195 not only allow the absorption of biologically active peptides through the rectal, vaginal or nasal mucous membranes, but also a pulmonary absorption.

By the invention it is possible to biologically active peptides, such as. As insulin, in aqueous or oily solutions for rapid and largely complete absorption via the respiratory tract bioavailable, without side effects, such as irritation of the mucous membranes and allergic reactions occur.

The surprising effect of the effect of EFK as absorption promoter for biologically active peptides in the case of absorption via the respiratory tract is described using the example of inhalational insulin administration in mice.

A rapid onset of blood glucose reduction was achieved, which in its time course corresponds to a subcutaneous injection of old insulin.

Proof of efficacy was obtained by enzymatic determination of blood glucose according to 2. AB of the GDR by means of commercial Fermognost blood sugar cutlery. The absorbance was determined at wavelength 530 nm versus blank. The

Blood collection was performed before and at 10-minute intervals after application from the retroorbital venous plexus.

The surprising effect of the EFK with regard to insulin uptake via the respiratory tract could be detected as concentration-dependent both on the EFK used and on the insulin.

For animal experiments a device according to WP A61D 2414545 was used for inhalation.

The inhalable preparation needed for the application was prepared by means of an ultrasonic generator so that 30 ml of liquid

within 30 min in a m ^{3 of} air were fogged.

As test animals, metabolically healthy, not anesthetized, female hybrid mice (F ₁ ), the inbred tribes NMRI / BC x C57B1 / BC were used with an average body mass of 24 g, which were pre-tested for insulin sensitivity. The animals were deprived of food 17 hours before the start of the experiment. They received water ad libitum.

Four experimental groups were formed.

1. Inhalation of 25mg insulin, which was dissolved in 30ml insulin dilution (2nd AB / DDR) containing 5% EFK.

2. Inhalation of 30 ml of physiological saline

3. Inhalation of 25mg insulin in 30ml insulin dilution (2nd AB / DDR) without EFK.

4. Inhalation of 30 ml of physiological saline after subcutaneous administration of 0.6 ng insulin / kg mouse

 According to the respiratory rate and the respiratory volume of the mouse, the amount of inhaled insulin corresponds to approx. 0.6-1.0 ng / kg mouse. The effect of sc administration of insulin was made comparable by subjecting the animals to the same experimental conditions.

Examples

Blood glucose values in% of the initial values

1. Inhalation of insulin + EFK 5%

Animal no. - 20 min

1 64.7

2 88.3

3 110.6

4 104.9

5 86.0

6 103,2

7 90.2

8 '101.9

9 81.0 10 95.6

40 min

60 minutes 105.9 86.3 76.6 150.1 114.0 110.8 121.2 107.8 104.8 126.7

80 min

78.4

76.4

104.2

164.4

128.0

132.4

163.6

98.0

88.1

108.9

92.7 13.6

Blood glucose values in% of the initial values

2. Inhalation of physiol. Saline

Animal no. 20 min

1 126.6

2 116,1

3 131.3

4 136.5

5 130.0

6 185.3

7 192.6

8 156.7

9 161.3 10 166.7

40 min

110.4 20.4

60 min 190.4 197.7 150.0 177.0 150.0 188.2 218.5 213.3 245.8 205.6

114.2 32.1

80 min

182.9

236.8

166.7

228.4

157.4

217.6

233.3

253.3

303.2

152.8

150.3 26.1

Blood glucose values in% of the initial values

3. Inhalation of insulin without EFK

Animal 20 min

1 125.4

2 115.3

3 129.3

4 '102,7

5 116.2

6 140.9

7 113.3

8 161.3

9 128.6 10 146.7

128.0 17.6

40 min

193.7 29.8

60 min 189.8 177.6 197.8 118.8 177.0 152.5 186.7 212.9 175.0 153.3

174.1 26.8

213.2 48.0

80 min

196.6

154.1

208.7

183.9

171.6

190.1

209.7 217.9 190.0

191.4 20.0

Blood glucose values in% of the initial values

4. Inhalation of physiological saline + sc administration of insulin

Animal no. 20 min 40 min

1 66.6 · 83.3

2 84.1 89.3

3 .89.9 125.0

4 86.5 109.2

5 65.3 82.7

6 70.7 94.2

-3- 261 096 60 min 80 min 104.1 108.3 84.1 120.9 139.9 150.0 113.7 159.2 91.4 104.5 108.3 125.0 106.9 128.0 19.5. 22.2

X 77.2 97.3

s 10,9 16.7

significance

The test for significance was carried out by t-test according to STUDEND.

It has been found that with an error probability of 5%, the EDF-containing insulin preparations differ significantly in their blood glucose lowering activity from the insulin preparations without absorption mediators. No significant difference could be detected between the inhalative application of insulin with the addition of EFK and the subcutaneous administration of insulin.

Claims (2)

1. A process for the preparation of inhalable resorbable peptide preparations, in particular of insulin, characterized in that the active ingredient is added to a surfactant. 2. The method according to item 1, characterized in that egg white fatty acid condensates are used as the surfactant. Field of application of the invention The invention relates to the production of biologically active substances having a peptide structure, which are brought to absorption in the form of suitable preparations in human and veterinary medicine via the respiratory tract.