JPS61183228A - Drug preparation for external use - Google Patents

Abstract

PURPOSE:To provide a drug preparation for external use, by adding negamycin or its salt to a substrate containing at least an aliphatic alcohol and a glycol solvent. CONSTITUTION:The objective drug preparation for external use can be produced by adding 0.5-3% negamycin or its salt to a substrate obtained by mixing (A) 15-45% (in the substrate) aliphatic alcohol (e.g. 16-24C saturated aliphatic alcohol such as cetyl alcohol, stearyl alcohol, etc.), (B) 45-85% (in the substrate) glycol solvent (e.g. propylene glycol such as 1, 2-propanediol or a polyethylene glycol having a molecular weight of 100-800) and if necessary (C) a plasticizer (e.g. polyethylene glycol having a molecular weight of 800-20,000), a binder (e.g. stearic acid) or a penetrant (e.g. dimethyl sulfoxide). The drug is effective for the remedy and prevention of infectious diseases caused by Gram-positive bacteria, Gram-negative bacteria or their resistant mutant.

Description

DETAILED DESCRIPTION OF THE INVENTION Technical Field of the Invention The present invention relates to an external preparation containing negamycin or a salt thereof as an active ingredient.

Prior Art Negamycin (hereinafter referred to as NGM) was discovered by the present inventors, and was isolated from Streptomyces Purpeofuscus.
It is an antibiotic that is produced by a few seconds of actinobacteria belonging to the U.S. species and has particularly excellent antibacterial activity against Gram-negative bacteria (
It is also known to be effective against plant diseases caused by bacteria (see Japanese Patent Laid-Open No. 1746-12925P3). Streptomycin (Str-1-1-11-mycin) causes misreading of the mRNA codon due to its mechanism of action.
ptomycinl, kanamycin (Kanamycin)
Although it is similar to aminoglue 1-cyto antibiotics such as (Y, 1Io11ara etal,
: J Antibiotics, 25.
865 (1972)), NGM is known to particularly strongly impair the termination process of protein synthesis (Y, 1lehara, H.
,1lori, 11. lImezr+wa: Bi
ochim.

Biophs, octa, 374. 82 (
1074) ; 442. 2!11 (197G
)),.

By the way, in connection with the daily use of antibiotic therapy, the transformation of infectious diseases in recent years has been dizzying, and the emergence of drug-resistant bacteria caused by Gram-negative bacteria, including B. aeruginosa, has been particularly dramatic41. There is one C assignment.

In addition, the induction of secondary infections or other infections from burns, trauma A3, postoperative wound surfaces, etc. caused by these bacteria has become a familiar problem for us.

N9M can be said to be a particularly effective antibiotic for the treatment and prevention of the above-mentioned infectious diseases, and is highly anticipated (Patent Application Fl. 46-28835A, J.
biotics, 23.170 (1970)), however, there are problems with safety and release properties in the base material, and it has not yet been put into practical use. This is the current situation.

① Overview 1 The present invention aims to solve the problem mentioned in ②.As a result of examining the release properties, safety and effectiveness of NGM in external preparations, the present invention has been developed to reduce the amount of aliphatic alcohol and glycol solvent. This is based on an unexpected discovery even for those skilled in the art: by using a base containing 1,000 ml of NGM, it is possible to bring NGM to a degree of sterilization in a short period of time without causing decomposition of the crude drug.

The external preparation according to the present invention is characterized by containing Negamycin J or a salt thereof in a base containing at least an aliphatic alcohol and a glycol solvent.

Treatment I In the external preparation of the present invention, a base containing at least an aliphatic alcohol and a glycol solvent is used. Increases release properties,
1. By being able to stably retain and coat NGM in a layer, it is possible to prevent various infections caused by Durum-positive bacteria, Durum-negative bacteria such as Tit aeruginosa, and their resistant bacteria (such as hazy rash, It is believed that this invention will make a significant contribution to the external treatment and prevention of cystitis, trichoentery vulgaris, superficial blepharoderma, secondary infections, etc.).

Furthermore, in addition to Appendix IJ, the present external preparation using the I-marker has the following advantages.

(b) It is a IL that can be widely applied to both cream-based and oily-based lesions.

([1) Can be easily washed with water.

(c) Adheres well to injured areas moistened by mucous membranes or secretions.

As mentioned above, NGM is a known substance, and the structure of A is δ-hydroxy-β-lysine (δ-hydroxy-β-lysine).
hydroxy-β-+ystne) and 1-methylhydrazino PA acid (1-methylhydrazino
acetic acid). In addition to the above-mentioned compounds, the NGM used in the present invention also includes any derivatives that exhibit antibacterial activity (Japanese Patent Application Laid-Open No. 52-1992).
No. 591'12 and Japanese Unexamined Patent Publication No. 52-83605). These compounds may also form salts with pharmaceutically acceptable inorganic or organic acids such as strong acids such as hydrochloric acid, sulfuric acid, nitric acid, benzoic acid, etc., and the present invention therefore also covers salts of NGM. It is.

NGM is Streptomyces purpeofusc.
several species of actinomycetes (M890-02 strain (Microbial Chemistry Institute strain number), M△91-, M1 strain (Microbial Chemistry Institute strain number), MAi04-MI strain (Microbial Chemistry Institute strain number) ), etc.), or synthetically (JP-A-52-59112, Yoji Ohno et al.: Journal of the Chemical Society of Japan, 9.1299) (1983)). Therefore, it is best to synthesize NGM as necessary or to obtain it by culturing the actinomycetes as described above.

NGM is generally a white powder, odorless, and soluble in water.Among lower alcohols, NGM is slightly soluble in methanol, but is sparingly soluble or insoluble in other organic solvents.
be.

NGM generally has low toxicity (1-D to mice).
5o is 9 or more in 200 m97 by intravenous injection (see JP-A-52-59112).■ The base used in the present invention contains at least an aliphatic alcohol and a glycol solvent, and if necessary Depending on the situation, a plasticizer, a binder, a penetrating agent, etc. are mixed therein. A specific example of this is the △PG antidote developed by Margin Carla et al.
(See March statement).

(a) Aliphatic alcohol The aliphatic alcohol used in the base material is preferably a higher-valent alcohol. Specifically, carbon number 16-24
Suitable saturated aliphatic alcohols include, for example, lebrual alcohol, stearyl alcohol or behenyl alcohol.

The concentration of these compounds in the base is usually 15-4.5%, preferably 20-35%.

(B) Glycol solvent The glycol solvent used in this base has two (or several) hydroxyl groups attached to a non-hydrocarbon M (approximately 2 to 4 carbon atoms) residue or a hydrocarbon containing an ether bond ( Grinding carbon number 4~
(about 36) residues are preferred. These are known to act as solvents for drugs that are soluble in glycol, or as carriers for drugs that are not soluble in glycol. 2
Copanediol, J3J, and 1,3-propanediol, as well as polyethylene glycol with a molecular weight of 100 to 800! - Le, Ji/[
] Pipylene glycocol or these substances.

The mark J in the base of these compounds is usually /15・he・B
! '1%, which is exactly (,1,55~ε30%
(·be.

The above compound ((a), J, bi(+”l)) is
The method of Martin Carla et al. (Kiguchi, 1 special h'1 No. 359
2930 S-3 specification f”) to J, S “Δp, (3
It can be prepared as a 1E drug.

4j The above ↓ agent may contain a plasticizer (molecular weight 80
0 to 20,000 poly]-tylene glycol, 1
．． 2.6-hekylyl-lyl, sorbyl-1-lyl, glyceryl-1-1-lyl, etc.), binders (stearic acid, palmitic acid, behenic acid, etc. with 16 carbon atoms and 2/I saturation) fatty acid,
Fatty acid amides such as Areamide, palmicamide, Sude) 7lamid, BeheJmid, sorbitan monosuide) Ru-1,
Glycerin tnoscyare-1-, zo[]]pyrenguly: 1-rumonosteare 1~ etc. to Ij2 prime number 16・2'
J, k may be mixed with a penetrating agent (dimebrusul small onitoside, dimethyl 77mide, dimebrus small lumamide, etc.).

External preparation The external preparation according to the present invention contains the essential ingredients of NGM and a base containing a small amount of aliphatic alcohol and glycol solvent, as well as auxiliary ingredients such as a plasticizer, a binder, a penetrating agent, and a preservative. ointment (
(including oral ointments and eye ointments), creams, and other formulations. If necessary, other drugs such as adrenal hormones (betamethacin, hydrocortisone, etc.), other antibiotics, or antihistamines (diphenhydramine, etc.) may be added.

A preferred formulation form of the external preparation of the present invention is a soft formulation.

The composition of this external preparation may be changed as appropriate depending on the dosage form, but
Usually, the wood agent contains 0.5% to 3% NGM.

The above-mentioned external preparations can be stored, for example, in an airtight container and tightly sealed, or in a bucove or the like.

Directions for use: The dosage is usually adjusted according to the symptoms.
To +1, apply it several times to the affected area (11'1), or spread it out and apply aseptically (-1).

The external preparation according to the present invention is effective in treating and preventing infections caused by Durham-positive bacteria, Togra 18-negative bacteria, and resistant bacteria.

41 The formulation used in this case has N G M /, + < low toxicity, and the base j3 and auxiliary ingredients are contained in the formulation, so there is no problem in terms of toxicity. Seem.

Experimental example 1) Quantification of NGM (a) High performance liquid lithography [171-graphy (++Plc
) in 0.05 M phosphate buffer (p 1-
16.8) with excessive premature fluorescamin (P, Boh
len, etal,, 8rcbBiochcm
, B10plT1/S, , 163.390 (197
4) Add the Dionisane solution of ) and stir with an electric mixer for about 1 minute to prepare the sample, and then perform high performance liquid chromatography (hereinafter referred to as 1-IPLC) (pump: 1655 in mouth, Kara lx: NLJC
Stainless steel column (8 mm x 250 ml++) packed with I
Internal standard substance: D-glutamic acid, mobile phase: methanol 10.1M acetic acid mixture (40:60)), and the eluted fluorescent substance was detected using a detector (1) Stand 650-10I.
C) with an excitation wavelength of 390 nm and a fluorescence wavelength of 47 nm.
'Measurements were made at 7 nm. N obtained in this way
GM and 71] Fluoresamine
The chromatography of the complex with is shown in FIG.

NGM and) lorelimine (fluorescamine)
) complex with retention time 9.1 min, 13.4 min a3
The retention time of the peak (5) of the internal standard substance (D-glutamic acid) was 28 minutes. ), select peak 4 with the best disintegration among them /υ (2 molecules of florerimin (
The relationship between the peak height ratio and the concentration of NGM is
A calibration curve (Fig. 2) was prepared and used in experiments below C1.

(1) Quantification of Pseudomonas aeruginori (“in cup γ”)
Saeruginosal I F O3923 was tested using 18 soils (11 water) for t'Lj-r screen (cups (8
φ x 10 mm) and 0.05 M phosphate buffer (pl
After filling the cup with NGM serially diluted with -16,8), the cup was incubated at 37°C for 19 hours, and the closed circle was 1 yen (m
m ) and create a standard curve (Figure 3).

As a result of (a), it was observed that the diameter of the field increased by 11 yen in proportion to the electrical production of NGM. Therefore, in subsequent experiments, N G
The increase or decrease in the release amount of M is determined by C judgment based on the stop ring diameter (mrn).

2) Stability of NGM I No 1 Test Experimental force method: NGM-containing (1.0 w/w%) ointment trial test approx. 2
g was sealed in a non-purple bottle and stored in a 25°C incubator. After 2 hours, remove the ointment sample and place it in a centrifuge tube (50 g
%) and 300 my each. After adding 5d of Roborum to the solution and dissolving the ointment sample, 10 parts of Flll-16,8 phosphate buffer was further added and the mixture was shaken vigorously for 30 minutes. Then, centrifugation #I (25
0 Or, p, 'm +' 10 m1n), sample the aqueous phase of the supernatant, and quantify it using the above-mentioned NOJ tube method. 1,0w/w%) M rounding diameter of soft husk/sample (
mm> was set to 100% to display the remaining amount of NGM).

B1. Each measurement was performed three times.

(A)...N GM (1, Ow/w%) containing oil-based base (Plastibase ■50 W (r, R, 5qui
bb&5ons) ) (b)...NGM (1,Ow/w%)-containing water-absorbing base (hydrophilic bo[''id) (Maruishi Pharmaceutical) (c)...NGM (1,0w/w%) Contained FΔPG
Base (Contains 30% stearyl alcohol, 0% propylene glycol) Experimental results
quibb & 5ons) in N GM (1,0
w/w%) and water absorption ↑su1 (agent hydrophilic point 1id■ (Maruishi Pharmaceutical)) and NGM (1,0w/w%) in FAPG base. The results of the stability comparison are shown in the table below.

NGM was stable in all three types of bases that did not contain water. Inside 5 Plastibase ■ 50W (lR
,5quihl)&Sor+s)OJ:biFAPG
It was stable at 1St in the base.

Note that NGM is unstable (・
It is also known that there is such a thing (see specification of Japanese Patent Application Laid-Open No. 1983-22983).

3) Experimental method for release of NGM from a famous ointment base. etal,,
CI+em, Pt+armBu11. ．． 6. 24
01.-2405 (1984)) (The septum was a 15.9 cm cellophane membrane (Saj
rius, Barrierefolie SM 16
75 II) and 0.05M phosphate buffer (t) l-16,8) as the release solution.
12ml! /min - (: Circulate the discharged liquid 1171]!
), ・NGM released over time in the above 1-11)
Fj4 was determined according to the quantitative method of

- Soft base / base - (A)...N GM (1,0 w/w%) containing oil base ■ (Plastibase 50 W (E, R, Sqt old)
g: 5ons) ) ([1)...NGM (1,0w/w%)-containing oleaginous base (Po'Ido■ (Maruishi Pharmaceutical)) (c)...NGM (1,0w/w %) containing water-absorbing base (hydrophilic bo[-lid (Maruishi Pharmaceutical)) (c)...NGM (1.0 w/w%) containing FAPG
The release behavior from the base (30% stearyl alcohol, 0% bu[]]pyrene glycol]-lua) is shown in Figure 4 of JL, and the release behavior from the FAPG base is shown in Figure 5. be.

For the release of NGM, F APG base is the most
Then, water-absorbing base (hydrophilic boroid (R''), oleaginous base (boloid ■, Plastibase ■50W) in the order C
there were.

4) "Experimental method for NGM release from △PG base"
Asaeruoinosa) I F O3923 was tested using a culture medium for sensitive disks (Hiinaga) to a test bacterial count of 1 x 106.
NGM was added to a thin layer plate prepared to have a thickness of 1, Qw/mQ.
A cup (φ8 x 10 mm) filled with ΔPG base with the following composition containing w% was placed and cultured at 37°C for about 191 hours, and the till ring diameter (mm) was measured (Ronsuke Sumiki et al., "Antibiotics (1)", University of Tokyo Press, Tokyo,
1.9'6'l, Pl 22>.

= 16 - - - F = A PG base - 1 Experimental results The release properties of NGM from F A PG bases of various compositions were as shown in the table below. Nao Tokikinari no Ura △-D
is a typical F A PG base.

As a result, NGM in the -L distribution U APG base (A to F) showed almost the same inhibition circle diameter as ()C1 compared to -B Sycode Mobbs Engineering Luginault 4 No. IF-03923. . Therefore, it has become clear that at least an aliphatic alcohol and a glycol solvent are required in the F APG base.

5) Experimental method for antibacterial activity of NGM using FAPGI agent Staphylococcus aureus (5 tapyl.

coccus aureus) IF O3060
, Pseudomonas eruginos 4 no.
nas aeruginosa) IFO3923, Proteus vulgaris (Proteus vulgari)
s) IFO3167 and Serratia marces-cens
ce) Four types of test bacteria of I FO3046 were placed on a 49-layer plate prepared using Mueller-Hinton medium (formerly FCO) to a test bacterial count of 1 x 106 cells/d. 1 each of M w/
w% containing "ΔPG base (stearin) Fluor 1-25%, stearic acid 5%, B-1-1 pyrene glycerin"-
A cup (φ8 x 10m) filled with
After culturing at 37°C for about 19 hours, the inhibition circle diameter (mar) was measured (Ronsuke Sumiki et al., 1 anti/1 substance (g), University of Tokyo Press, Tokyo Tech; 1961, P.I.
22>.

M-ru N0M content (1.0 w/w%) F for various test gold at the left end of actual J eye
APG Soft Blue Goods 111 - The diameter of the circle is as shown in the table below.

As a result, the 111L circle diameter showed the most popular size for Cycodermovus luginori Il-03923J5 and Proteus vulgaris IFO3167, followed by Rl7a marcellens IFO3,046.
, Staphylococcus arale [us IFO3060].

1 20 -

[Brief explanation of the drawing]

Figure 1 shows chroma 1 of NGM and florerimin complex.
This is a copy of the Hegraph Society. FIG. 2 is a diagram showing a calibration curve showing the relationship between m degrees of NGM and chroma I to peak height ratio of the graph. Figure 3 shows the NGM (7) concentration and Sycordomonas.
It is a figure which shows the standard curve which showed the stop diameter of NGM with respect to Elino-4. FIG. 4 is a graph showing the amount of NGM released in the oily base (Plusdibase ■5°W and 0 bolloid), J2 and the hygroscopic base (hydrophilic bolloid ■) over time. FIG. 5 is a graph showing the amount of NGM released from [ΔPG base] over time. Applicant's representative: Inomata Kiyoji Time (minutes) Figure 2

Claims (1)

[Claims] An external preparation comprising negamycin or a salt thereof in a base containing at least an aliphatic alcohol and a glycol solvent.