JPS62149623A - Stable growth hormone releasing factor (grf) pharmaceutical - Google Patents

Abstract

PURPOSE:The titled pharmaceutical containing a human blood serum albumin or glycine and, as necessary, a buffer agent. CONSTITUTION:A stable growth hormone releasing factor (GRF) pharmaceutical obtained by adding 100mug-30mg, based on 100mug GRF (a peptide expressed by the formula), human blood serum albumin or glycine as a stabilizer to the GRF and further adding a buffer agent, e.g. acetate, phosphate, citrate, etc., as necessary, to give 2-7pH, preferably 2.5-5pH for providing a stabler pharmaceutical. The dosage form may be either one of solution of solid form, but is preferably the form of freeze-dried product as a medicine stable for a long period. In this case, nitrogen is more preferably sealed after freeze-drying. Furthermore, the pharmaceutical is readily dissolved by adding a suitable dissolvent solution in use and particularly useful as a simple freeze-dried injection of the type soluble in use.

Description

DETAILED DESCRIPTION OF THE INVENTION The present invention relates to stable GRF formulations. More specifically, the present invention relates to a GRFII agent characterized by containing human serum albumin or glycine as a stabilizer.

GRF (Growth Hormone Releasing Factor)
) is a bebutide purified and isolated from the pancreas of a barrel tumor patient with symptoms of acromegaly by Guillemin et al., and has the following amino acid-substructure: H-'ryr-Ala. -Asp-Ala-11
e-Phe-Thr-Asn-3er-Tyr-Arg
-Lys-Val -Leu-Gly-Gln-Leu
-5er-Ala-Arg-Lys -Leu-Leu
-Gin-Asp-I 1e-Me t-3e r-A
rg-Gln-Gln-Gly-Glu-5er-As
n-Gln-Glu-Ar g-Gly-A 1
a-Ar g-A l a-A r g-L e
u-NHZ [Science x:/Science, μ
8, 585-587° It was subsequently shown to be structurally identical to GRF purified from the lower part of a felled tree, and GRF (7) was replaced by somatocrinin (Somatocrinin).
The name ocrinin) has been proposed.

GRF has so far had amino acids 44, 40, 37 and 2.
Each of these peptides has been separated and purified as a peptide consisting of 9, and it has been confirmed that all of them exhibit GH (Growth) (ormone) release activity. These peptides are expected to be administered temporarily or long-term for diagnostic or therapeutic purposes in clinical medicine (both human and veterinary medicine).

However, GRF does not have sufficient storage stability in solution, so when developing it as a drug, it is desirable to make it into a lyophilized formulation, but even with lyophilization, its stability is not always sufficient (at room temperature A significant decrease in titer is observed under long-term storage, heating, humidification, or light conditions.

It is known that one of the causes of this is that the methionine at position 27 of the 171 amino acids constituting GRF is easily oxidized. Therefore, a stabilization method of adding an antioxidant such as avic acid (no change in iI+minus content of L-Ascol light #l*) was investigated, but it was still not possible to obtain a freeze-dried preparation that was stable for a long period of time.

As a result of intensive studies aimed at improving these points, the present inventors discovered that by adding human serum albumin or glycine, a stable and practically extremely useful GRF preparation could be obtained, and the present invention was completed. This is what I did. The GRF in the GRF preparation of the present invention may be any peptide that exhibits growth hormone-releasing activity; for example, it may have any number of amino acids ranging from 44° to 40.87.29, or may be a mixture thereof. In addition, the present invention G
There are no particular restrictions on the shape of the RF preparation; for example, it may be in the form of a solution or a solid, but as a long-term stable drug, it is desirable to have it in the form of a lyophilized product. It is more desirable to fill it with nitrogen after drying.

The lyophilized preparation of GRF of the present invention not only has remarkable stability compared to preparations containing other additives, but also can be prepared by adding a suitable solution for injection, such as distilled water for injection, physiological saline, etc. It has the advantage of being easily soluble, and is particularly useful as a simple injection-dissolving type freeze-dried injection.

Here, the amount of human serum albumin or glycine added is
Although not particularly limited, 100 μy to 30 cape is suitable for GRF 100 μm.

In addition to human serum albumin or glycine, a buffer may be added as necessary to obtain a more stable preparation. Examples include acetate, phosphate, citrate, etc. The amount of buffering agent to be added is, for example, in the case of liquid preparations such as injections and nasal drops, when the pH of the preparation is 2 to 7, preferably 2.5 to 5, considering the physical and chemical stability of GRF. Just add it so that In the case of a freeze-dried preparation, it may be added so that the pH of the preparation after redissolution becomes the above value.

It goes without saying that the preparation of the present invention may also contain conventional additives such as isotonic agents, soothing agents, excipients, and the like. The preparation of the present invention may be produced by a conventional method, but for example, when producing an injection, add macroserum albumin or glycine to a syringe of distilled water or a suitable mild liquid system, dissolve GRF, Next, sterile filtration may be performed. This is further freeze-dried using the method described in 1.at.

The present invention will be explained in detail below using experimental examples.

Experimental Example-1 The following items were investigated to confirm the stabilizing effect of the present invention. As a result, human blood was confirmed.

Experimental Example 2 An experiment was conducted to confirm the stabilizing effect of the present invention on GRF (1-29). Various stabilizers were dissolved in a phosphate buffer solution in which GRF (1-29) was dissolved, and then freeze-dried and sealed with nitrogen, and the items shown in Table 2 were investigated.

As a result, it was found that the freeze-dried preparation to which human serum albumin or glycine was added had remarkable stability compared to the case without the addition of human serum albumin or glycine. Furthermore, in the freeze-dried preparation containing cysteine and sodium thiosulfate, the solubility deteriorated and the stability significantly decreased.

Examples are shown below, but it goes without saying that the present invention is not limited thereto.

Example 1 Human serum albumin 20011y was added to 10 g/l of acetic acid-sodium acetate buffer at pH 5, and then GRF (
1-44) 10q was dissolved. After sterile filtration of this solution,
00μβ was filled into vials and freeze-dried. After filling with nitrogen, a rubber stopper was placed and the cap was sealed to obtain a stable lyophilized injection.

Example 2 Human serum albumin 100114 was added to 50 yrl of PH3 phosphate-citrate buffer, then GRF (1-
44) Dissolved 10 q. After sterile filtration of this solution,
0 μl each was filled into vials and freeze-dried. After filling with nitrogen, a rubber stopper was placed and the cap was sealed to obtain a stable lyophilized injection.

Example 3 Glycine 400''5' was dissolved in 50 g of distilled water for injection, and then 10 μg of GRF (1-44) was dissolved. After sterile filtration, this solution was filled into vials in 500 μβ portions and freeze-dried. After filling with nitrogen, a rubber stopper was placed and the cap was sealed to obtain a stable lyophilized injection.

Example 4 GRF (1-29) 4■ at 1/10 M I of pH 5
J acid buffer 20mj! Human serum albumin 4 after dissolving in
0 ■ was added. After sterile filtration, this solution was filled into vials at 5aOttl portions and freeze-dried. After filling with nitrogen, a rubber stopper was placed and the cap was sealed to obtain a stable lyophilized preparation.

Example 5 After dissolving 4 ml of GRF (1-29) in 20 ml of 1/10M phosphate buffer at pH 5, 40 ml of human serum albumin and 280 ml of sodium chloride were added. After sterile filtration, this solution was filled into vials with 500 μβ portions and freeze-dried. After filling with nitrogen, a rubber stopper was placed and the cap was sealed to obtain a stable lyophilized preparation.

Example 6 GRF (1-29) 4■ with 1/10M phosphoric acid at pH 5! ! After dissolving in 20 ml of street liquor, 40 ml of human serum albumin and 360 ml of sodium chloride were added. After sterile filtration, 500 μl of this solution was filled into vials and freeze-dried. After filling with nitrogen, a rubber stopper was placed and the cap was sealed to obtain a stable lyophilized preparation.

Example 7 After dissolving GRF (1-29) 4■ in 1/10M phosphate buffer 2 QrnJ of pH 5, human serum albumin 4 (1
+t and 400 μl of Funnitol were added. After sterile filtration, 500 ul of this solution was filled into vials and freeze-dried.

After filling with nitrogen, a rubber stopper was placed and the cap was sealed to obtain a stable lyophilized preparation.

Example 8 GRF (1-29) 20■ at 1/10 M of pH 5
Human serum Fulpmin A after dissolving in 20ml of IJ acid buffer
nw apricot was added. After sterile filtration, 500 μl of this solution was filled into vials and freeze-dried. After filling with nitrogen, a rubber stopper was placed and the cap was sealed to obtain a stable lyophilized preparation.

Example 9 GRF (1-29) 20aw with 20mj of l/10M phosphate buffer at pH 5! After dissolving in human blood albumin 2
00 ■ and 280 ■ of sodium chloride were added. After sterile filtration, 500 pl of this solution was filled into vials and freeze-dried. After filling with nitrogen, a rubber stopper was placed and the cap was sealed to obtain a stable lyophilized preparation.

Example 10 After dissolving 20 ml of GRF (1-29) in 100 ml of 1/10M phosphate buffer at pH 5, human serum albumin 200
Qr and 1.4 g of sodium chloride were added. After sterile filtration, this solution was filled into vials in 2.5 mjt portions and freeze-dried. After filling with nitrogen, a rubber stopper was placed and the cap was sealed to obtain a stable lyophilized preparation.

Claims (5)

[Claims] (1) GRF preparation containing human serum albumin or glycine and, if necessary, a buffering agent (2) The buffering agent is a buffering agent that maintains the pH value of the preparation at about 2 to 7 by adding it.
GRF preparations described in section (3) The GRF preparation according to claim 1, wherein the preparation is in the form of a lyophilized product. (4) The GRF preparation according to claim 3, wherein the buffering agent is a buffering agent that maintains the pH value of the preparation after redissolution at about 2 to 7 by adding it. (5) The GRF preparation according to claim 3 or 4, wherein the form of the preparation is a freeze-dried product filled with nitrogen.