CA1188615A - Stable nonaqueous solution of tetracycline salt - Google Patents
Stable nonaqueous solution of tetracycline saltInfo
- Publication number
- CA1188615A CA1188615A CA000384103A CA384103A CA1188615A CA 1188615 A CA1188615 A CA 1188615A CA 000384103 A CA000384103 A CA 000384103A CA 384103 A CA384103 A CA 384103A CA 1188615 A CA1188615 A CA 1188615A
- Authority
- CA
- Canada
- Prior art keywords
- percent
- nonaqueous
- tetracycline
- solution
- solubilizer
- 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
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K9/00—Medicinal preparations characterised by special physical form
- A61K9/0012—Galenical forms characterised by the site of application
- A61K9/0014—Skin, i.e. galenical aspects of topical compositions
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K31/00—Medicinal preparations containing organic active ingredients
- A61K31/65—Tetracyclines
Abstract
ABSTRACT OF THE DISCLOSURE
Nonaqueous solution of a tetracycline antibiotic salt which is stable upon extended storage comprises a mixture of a tetracycline antibiotic salt, nonaqueous diluent, nonaqueous solvent, and nonaqueous nonionic solubilizer. It preferably also contains an anti-oxidant and a nonaqueous anionic solubilizer.
Nonaqueous solution of a tetracycline antibiotic salt which is stable upon extended storage comprises a mixture of a tetracycline antibiotic salt, nonaqueous diluent, nonaqueous solvent, and nonaqueous nonionic solubilizer. It preferably also contains an anti-oxidant and a nonaqueous anionic solubilizer.
Description
STABLE NONAQUEOUS SOLUTIO~
OF TETRACYCLINE SALT
BACKGROUND AND PRIOR ART
Tetracycline antibiotics and their salts are well~known therapeutic materials. It has been pro-posed in the prior art to employ solutions of such ~0 tetracycline antibiotics as topical therapeutics for the treatment of various disorders, such as acne vul-garis. Tetracycline antibiotics are known to degrade to form epitetracycline, anhydrotetracycline, epi-anhydrotetracycline and other degradation products 9 ~5 some of which are unidentifiedO These degradation products have negligible therapeutic activity. This degradation appears to increase when solutions of tetracycline antibiotics are employed. Solutions of neutral tetracyclines in nonaqueous solvents are dis-closed in U.S. Patents Nos. 3,219,529 and 3,389,174.These references, however, do not disclose or suggest stable nonaqueous solutions of tetracycline anti-biotic salts. In fact, these references specifically MS-1081-CIP ~
1 state that they are not concerned with such salts.
An aqueous ethanol solution of tetracycline hydro-chloride has been commercially marketed under the tradename "Topicycline'1, but it is relatively un-stable in its solution Eorm.
There is thus a need Eor a stable nonaqueous so-lution of tetracycline antibiotic salts.
SUMMARY OF THE INVENTION
In accordance with the present invention, a I0 stable nonaqueous solution of a tetracycline anti-biotic salt is provided comprising a mixture of a tetracycline antibiotic salt, nonaqueous diluent, non-aqueous solvent and nonaqueous nonionic solubili7er.
Preferably, the solution also contains an antioxidant and a nonaqueous anionic solubilizer.
DESCRIPTION OF THE INVENTION
The tetracycline antibiotic salts used in the present invention are well-known and are readily available from several commercial sources. These salts are, for example, tetracycline hydrochloride, chlor-tetracycline hydrochloride, oxytetracycline hydro-chloride, rolitetracycline hydrochloride, and the like. Tetracycline hydrochloride is the preferred active ingredient.
The nonaqueous solvent can be ethanol, methanol 9 isopropanol, butanol, N-methyl-2-pyrrolidone or ace-tone. Ethanol is the preferred solvent.
The nonaqueous diluent which serves as the carrier for the overall composition can be an ester, 1 amide, silicone, ~ri~lyceride and ~he like. Suitable esters include glyceryl triaceta~e, diisopropyl sebacate, diisopropyl adipate 9 isopropyl palmitate, isopropyl myristate~ lauryl lac~ate, linear alcohol lactate, decyl oleate 9 isodecyl oleate, 2-ethylhexyl palmitate, isopropyl linoleate, acetylated mono-glyceride, acetyl tributyl citrate, acetyl triethyl citrate, tricyclo hexyl ci~rate, butyl myristate, promyristyl propionate~ butyl myristate, dibutyl 10 sebacate, dioctyl adipate, dioctyl succinate, and isobutyl ace~ate. Suitable amides include acetamide and methy]acetamide. Sui~able silicones include polydimethyl cyclosiloxane and hexamethyl disiloxane.
Suitable triglycerides include fractiona~ed tri-15 glycerides of coconut oil origin such as ~'Neobee M-5"
and "Neobee O" (Drew Chemical Corp.)O
- The nonaqueous solu~ilizers can be selected from two chemical groupings of materials, ~he nonionic sur-factant type materials and the anionic suractant 20 type materials. The nonionic surfactant type ma-terials preferably are of a mol. wt. range of 190 to 10,000 and can include polyethylene glycols, methoxy polyethylene glycols, polysorbates~ ethylene oxide-propylene oxide block copolymers, sorbit~n esters and 25 glycerin. The anionic surfactant type materials can consist of the monoYalent or di~alent salts of di-alkyl sul$osuccinates, acyl lactates and aryl sulfo-~ates. The pre~erred solu~ilizers consist of a mix-ture of a nonionic surfactant type material and an 30 anionic surfactant type mate~ial. The most pre-ferred solubilizers consist of a mixture of poly-ethylene glycol 200 and dioctyl sodium sul$osuccinate.
* Trade Mark ~ "' ' 1 The antioxidant or antioxidant combination in-cluded in this stable solution can consist oE ascorbyl palmitate, ascorbic acid~ propyl gallate, butylated hydroxytoluene, butylated hydroxyanisole, tertiary butyl hydroquinone, and d-beta, gamma and delta iso-mers of tocopherol as well as most other commonly known antioxidants .
The stable solution of the present invention gen-erally comprises a mixture of 0.25 to 2 percent tetra-10 cycline antibiotic salt, 41 to 88 percent nonaqueousdiluent, 4 to 40 percent nonaqueous solvent, 0.3 to 20 percent nonaqueous nonionic solubilizer, 0 to 6 percent nonaqueous anionic solubilizer and 0 to 0.75 percent antioxidant. These percent values are on a 15 weight/weight basis based on the entire solution weight.
A more preferred stable solution of the present invention comprises a mixture of 0.3 to 1.5 percent tetracycline antibiotic salt, 57 to 84 percent non-20 aqueous diluent, 4 to 30 percent nonaqueous solvent,2 to 9 percent nonaqueous nonionic solubilizer, 1 to 6 percent nonaqueous anionic solubilizer and 0.03 to 0.2 percent antioxidant.
The most preferred stable solution of the present 25 invention comprises a mixture of 0.5 t 1.25 percent tetracycline antibiotic salt, 60 to 81 percent non-aqueous diluent, 4 to 25 percent nonaqueous solvent, 3 to 9 percent nonaqueous nonionic solubilizer, 1.5 to 4.5 percent nonaqueous anionic solubilizer and 0.1 30 to 0.2 percent antioxidant.
The solution is conveniently prepared by mixing the ingredients in the following order. The non-aqueous nonionic solubilizer and the nonaqueou$
s 1 anionic solubilizer, if employed, are added to the nonaqueous solvent at ambient room temperature (about 20-25 C~), and the resulting mix-ture is stirred until disso]ved. Ihe tetracycline antibiotic sal-t and the antioxidant, if employed, are then added with mixing. The nonaqueous diluent is finally added.
The solutions of the present invention have been stored ~or extended periods of time at 22 C., 40 C., 50 C. and 60 C. The invention allows the tetra-10 cycline antibiotic salt to spontaneously epimerizeto an equilibrium condition assumed in the prior art to be about a 1:1 weight ratio of tetracycline to epitetracycline. This ratio is maintained and fur-ther tetracycline degrada-tion is inhibited by the 15 overall con~position so as to provide a therapeutic effective dosage of tetracycline antibiotic salt for an extended period of time.
The invention is described in further detail in the following examples.
EXAM~LE
A mixture was prepared having the following composition (Sample A):
s 1 Percent (weight/
weight basis) Tetracycline Hydrochloride 1.0 Dioctyl Sodium Sulfosuccinate 4.0 5 Polyethylene Glycol 200 4~0 Ethanol, 200 Proof 24.4 Glyceryl Triacetate 22.0 Diisopropyl Sebacate ~4.4 Ascorbyl Palmitate 0.2 A control composition (Sample B) consisting of an aqueous solution of tetracycline salt similar to that employed in the prior art was prepared having the following composition:
Percent (weightl weight basis) Tetracycline Hydrochloride 1.0 Ethanol, 200 Proof 40.0 Sodium Bisulfite 0.1 20 Water 58.9 100 . O
Appropriate samples of the above two compositions all having an input concentration of 10 mg./ml. tetra-cycline hydrochloride were stored at 22 C., 40 C., 25 and 50 C. for extended periods. The samples were periodically assayed for tetracycline, epitetracycline and other degradation products. The results are shown in the following Table 1. Though the formulations were initially prepared based on weight/weight per-30 centage amounts of the ingredients, their assay re-sults were reported as weight/volume percentages for convenience. The specific gravities of the compo-sitions were invariably close to 1.000 and any de-yiation therefrom introduces only errors of no sig-35 nificance into the results.
. ~
~1~ LO Lr~
~ ~ ~ ~ ~ o , V ~o ~ 1 00 ~ a~ oC~i C~l ~ C~
o ,.~ ~ 1 o ~ ~ ~ o ~ oo . ' _ ~ ~ O~ O C~ C~ CD O CD
~, o ~ ~ ""~ , lo bD ~ C~ o o o o - V V ~ ~ ~ ~ o o o~ o .
co ~ a) Lf~
o ~ ~ o o c~ c~ o a~ o c~ r-~ U2 0~ ~ L~ OO OCY~ C~ ~ O
o ~
~ 'r Lt' ~ Ll~ C~O
a~ ~ CD tD C~ ~ ~ cr~ c~ CO
~ ~C~) o o o o o ~ I ~ ~ ~
~ H V LO ~ Lf) OO OC~ d~ ~0 0 Cl ~ .
~1 ~ ¢¢ c~ r~41 ~ ~ o~ a~ o a~
~ c~ ~ o o o ~ ~
¢ ~ c~ L~Lr~ o Pll~L~ i~ ' E~ u~
~1 olo~ I O ~ olo ' lo ~ ~ ~ .
O s~ o O o ~, ~ r 9 ~ a V ~ i O
O L~ O
~ L~
1 Evaluation of the above data must take into con-sideration the previously mentioned epimerization re-ac-tion of tetracycline antibiotic salts wherein an e~uilibriu~n weight ratio of about 1:1 tetracycline to epitetracycline is spontaneously established in so-lutions of the salt. Thus for a stable target concen-tration of say 5 mg./ml., one must have an input level oE tetracycline hydrochloride of 10 mg./ml.
Relative stability thereEore of I0 mg./ml.
10 input formulas should be calculated as a comparison o-f the found value versus 5 mg./ml. of tetracycline hydrochloride. The composition oE the present in-vention ~Sample A) of Example l shows a retention of:
5~01 x lO0 = 100% tetracycline hydrochloride ]5 at 26 wks. 22 C.
4.65 x 100 = 93% tetracycline hydrochloride at 52 wks. 22 C.
4-99 x 100 = 100% tetracycline hydrochloride at 8 wks. 40 C.
4-85 x 100 = 97% tetracycline hydrochloride at 13 wks. 40 C.
-4 77 x 100 = 95% tetracycline hydrochloride 5 at 4 wks. 50 C.
4-57 x 100 = 87% tetracycline hydrochloride at 8 wks, 50 C.
Comparatively the control composition o~ the prior art aqueous solution ~Sample B) has a retention of:
- ;
s 1 4-$6 x 100 = 83% tetracycline hydrochloride at 26 wks. 22 C.
3-457 x 100 = 69% tetracycline hydrochlori.de at 52 wks. 22 C.
3-08 x 100 = 62% tetracycline hydrochloride at 8 wks. 40 C.
OF TETRACYCLINE SALT
BACKGROUND AND PRIOR ART
Tetracycline antibiotics and their salts are well~known therapeutic materials. It has been pro-posed in the prior art to employ solutions of such ~0 tetracycline antibiotics as topical therapeutics for the treatment of various disorders, such as acne vul-garis. Tetracycline antibiotics are known to degrade to form epitetracycline, anhydrotetracycline, epi-anhydrotetracycline and other degradation products 9 ~5 some of which are unidentifiedO These degradation products have negligible therapeutic activity. This degradation appears to increase when solutions of tetracycline antibiotics are employed. Solutions of neutral tetracyclines in nonaqueous solvents are dis-closed in U.S. Patents Nos. 3,219,529 and 3,389,174.These references, however, do not disclose or suggest stable nonaqueous solutions of tetracycline anti-biotic salts. In fact, these references specifically MS-1081-CIP ~
1 state that they are not concerned with such salts.
An aqueous ethanol solution of tetracycline hydro-chloride has been commercially marketed under the tradename "Topicycline'1, but it is relatively un-stable in its solution Eorm.
There is thus a need Eor a stable nonaqueous so-lution of tetracycline antibiotic salts.
SUMMARY OF THE INVENTION
In accordance with the present invention, a I0 stable nonaqueous solution of a tetracycline anti-biotic salt is provided comprising a mixture of a tetracycline antibiotic salt, nonaqueous diluent, non-aqueous solvent and nonaqueous nonionic solubili7er.
Preferably, the solution also contains an antioxidant and a nonaqueous anionic solubilizer.
DESCRIPTION OF THE INVENTION
The tetracycline antibiotic salts used in the present invention are well-known and are readily available from several commercial sources. These salts are, for example, tetracycline hydrochloride, chlor-tetracycline hydrochloride, oxytetracycline hydro-chloride, rolitetracycline hydrochloride, and the like. Tetracycline hydrochloride is the preferred active ingredient.
The nonaqueous solvent can be ethanol, methanol 9 isopropanol, butanol, N-methyl-2-pyrrolidone or ace-tone. Ethanol is the preferred solvent.
The nonaqueous diluent which serves as the carrier for the overall composition can be an ester, 1 amide, silicone, ~ri~lyceride and ~he like. Suitable esters include glyceryl triaceta~e, diisopropyl sebacate, diisopropyl adipate 9 isopropyl palmitate, isopropyl myristate~ lauryl lac~ate, linear alcohol lactate, decyl oleate 9 isodecyl oleate, 2-ethylhexyl palmitate, isopropyl linoleate, acetylated mono-glyceride, acetyl tributyl citrate, acetyl triethyl citrate, tricyclo hexyl ci~rate, butyl myristate, promyristyl propionate~ butyl myristate, dibutyl 10 sebacate, dioctyl adipate, dioctyl succinate, and isobutyl ace~ate. Suitable amides include acetamide and methy]acetamide. Sui~able silicones include polydimethyl cyclosiloxane and hexamethyl disiloxane.
Suitable triglycerides include fractiona~ed tri-15 glycerides of coconut oil origin such as ~'Neobee M-5"
and "Neobee O" (Drew Chemical Corp.)O
- The nonaqueous solu~ilizers can be selected from two chemical groupings of materials, ~he nonionic sur-factant type materials and the anionic suractant 20 type materials. The nonionic surfactant type ma-terials preferably are of a mol. wt. range of 190 to 10,000 and can include polyethylene glycols, methoxy polyethylene glycols, polysorbates~ ethylene oxide-propylene oxide block copolymers, sorbit~n esters and 25 glycerin. The anionic surfactant type materials can consist of the monoYalent or di~alent salts of di-alkyl sul$osuccinates, acyl lactates and aryl sulfo-~ates. The pre~erred solu~ilizers consist of a mix-ture of a nonionic surfactant type material and an 30 anionic surfactant type mate~ial. The most pre-ferred solubilizers consist of a mixture of poly-ethylene glycol 200 and dioctyl sodium sul$osuccinate.
* Trade Mark ~ "' ' 1 The antioxidant or antioxidant combination in-cluded in this stable solution can consist oE ascorbyl palmitate, ascorbic acid~ propyl gallate, butylated hydroxytoluene, butylated hydroxyanisole, tertiary butyl hydroquinone, and d-beta, gamma and delta iso-mers of tocopherol as well as most other commonly known antioxidants .
The stable solution of the present invention gen-erally comprises a mixture of 0.25 to 2 percent tetra-10 cycline antibiotic salt, 41 to 88 percent nonaqueousdiluent, 4 to 40 percent nonaqueous solvent, 0.3 to 20 percent nonaqueous nonionic solubilizer, 0 to 6 percent nonaqueous anionic solubilizer and 0 to 0.75 percent antioxidant. These percent values are on a 15 weight/weight basis based on the entire solution weight.
A more preferred stable solution of the present invention comprises a mixture of 0.3 to 1.5 percent tetracycline antibiotic salt, 57 to 84 percent non-20 aqueous diluent, 4 to 30 percent nonaqueous solvent,2 to 9 percent nonaqueous nonionic solubilizer, 1 to 6 percent nonaqueous anionic solubilizer and 0.03 to 0.2 percent antioxidant.
The most preferred stable solution of the present 25 invention comprises a mixture of 0.5 t 1.25 percent tetracycline antibiotic salt, 60 to 81 percent non-aqueous diluent, 4 to 25 percent nonaqueous solvent, 3 to 9 percent nonaqueous nonionic solubilizer, 1.5 to 4.5 percent nonaqueous anionic solubilizer and 0.1 30 to 0.2 percent antioxidant.
The solution is conveniently prepared by mixing the ingredients in the following order. The non-aqueous nonionic solubilizer and the nonaqueou$
s 1 anionic solubilizer, if employed, are added to the nonaqueous solvent at ambient room temperature (about 20-25 C~), and the resulting mix-ture is stirred until disso]ved. Ihe tetracycline antibiotic sal-t and the antioxidant, if employed, are then added with mixing. The nonaqueous diluent is finally added.
The solutions of the present invention have been stored ~or extended periods of time at 22 C., 40 C., 50 C. and 60 C. The invention allows the tetra-10 cycline antibiotic salt to spontaneously epimerizeto an equilibrium condition assumed in the prior art to be about a 1:1 weight ratio of tetracycline to epitetracycline. This ratio is maintained and fur-ther tetracycline degrada-tion is inhibited by the 15 overall con~position so as to provide a therapeutic effective dosage of tetracycline antibiotic salt for an extended period of time.
The invention is described in further detail in the following examples.
EXAM~LE
A mixture was prepared having the following composition (Sample A):
s 1 Percent (weight/
weight basis) Tetracycline Hydrochloride 1.0 Dioctyl Sodium Sulfosuccinate 4.0 5 Polyethylene Glycol 200 4~0 Ethanol, 200 Proof 24.4 Glyceryl Triacetate 22.0 Diisopropyl Sebacate ~4.4 Ascorbyl Palmitate 0.2 A control composition (Sample B) consisting of an aqueous solution of tetracycline salt similar to that employed in the prior art was prepared having the following composition:
Percent (weightl weight basis) Tetracycline Hydrochloride 1.0 Ethanol, 200 Proof 40.0 Sodium Bisulfite 0.1 20 Water 58.9 100 . O
Appropriate samples of the above two compositions all having an input concentration of 10 mg./ml. tetra-cycline hydrochloride were stored at 22 C., 40 C., 25 and 50 C. for extended periods. The samples were periodically assayed for tetracycline, epitetracycline and other degradation products. The results are shown in the following Table 1. Though the formulations were initially prepared based on weight/weight per-30 centage amounts of the ingredients, their assay re-sults were reported as weight/volume percentages for convenience. The specific gravities of the compo-sitions were invariably close to 1.000 and any de-yiation therefrom introduces only errors of no sig-35 nificance into the results.
. ~
~1~ LO Lr~
~ ~ ~ ~ ~ o , V ~o ~ 1 00 ~ a~ oC~i C~l ~ C~
o ,.~ ~ 1 o ~ ~ ~ o ~ oo . ' _ ~ ~ O~ O C~ C~ CD O CD
~, o ~ ~ ""~ , lo bD ~ C~ o o o o - V V ~ ~ ~ ~ o o o~ o .
co ~ a) Lf~
o ~ ~ o o c~ c~ o a~ o c~ r-~ U2 0~ ~ L~ OO OCY~ C~ ~ O
o ~
~ 'r Lt' ~ Ll~ C~O
a~ ~ CD tD C~ ~ ~ cr~ c~ CO
~ ~C~) o o o o o ~ I ~ ~ ~
~ H V LO ~ Lf) OO OC~ d~ ~0 0 Cl ~ .
~1 ~ ¢¢ c~ r~41 ~ ~ o~ a~ o a~
~ c~ ~ o o o ~ ~
¢ ~ c~ L~Lr~ o Pll~L~ i~ ' E~ u~
~1 olo~ I O ~ olo ' lo ~ ~ ~ .
O s~ o O o ~, ~ r 9 ~ a V ~ i O
O L~ O
~ L~
1 Evaluation of the above data must take into con-sideration the previously mentioned epimerization re-ac-tion of tetracycline antibiotic salts wherein an e~uilibriu~n weight ratio of about 1:1 tetracycline to epitetracycline is spontaneously established in so-lutions of the salt. Thus for a stable target concen-tration of say 5 mg./ml., one must have an input level oE tetracycline hydrochloride of 10 mg./ml.
Relative stability thereEore of I0 mg./ml.
10 input formulas should be calculated as a comparison o-f the found value versus 5 mg./ml. of tetracycline hydrochloride. The composition oE the present in-vention ~Sample A) of Example l shows a retention of:
5~01 x lO0 = 100% tetracycline hydrochloride ]5 at 26 wks. 22 C.
4.65 x 100 = 93% tetracycline hydrochloride at 52 wks. 22 C.
4-99 x 100 = 100% tetracycline hydrochloride at 8 wks. 40 C.
4-85 x 100 = 97% tetracycline hydrochloride at 13 wks. 40 C.
-4 77 x 100 = 95% tetracycline hydrochloride 5 at 4 wks. 50 C.
4-57 x 100 = 87% tetracycline hydrochloride at 8 wks, 50 C.
Comparatively the control composition o~ the prior art aqueous solution ~Sample B) has a retention of:
- ;
s 1 4-$6 x 100 = 83% tetracycline hydrochloride at 26 wks. 22 C.
3-457 x 100 = 69% tetracycline hydrochlori.de at 52 wks. 22 C.
3-08 x 100 = 62% tetracycline hydrochloride at 8 wks. 40 C.
2.79 x 100 = 56% tetracycline hydrochlori.de at 4 wks. 50, C.
- x 100 = 0% tetracycline hydrochloride at 8 wks. 50 C.
It can thus be seen from the above data that the composition of the present invention is considerably more stable on extended storage than a typical com-parable prior art composition.
Additional examples of varying compositions within the scope of the present invention are listed in the following Table 2. In addition, chemical stability data on a number of these formulations are given in Table 3.
-L~ o ~ O ~ O o O
C~ OtD C~J(:0r-l C~ O r--l ~ CO O
a~ c~ O o~
O o o oCl~ O ~ O
o C~ C~ ~ o ' o O
X ~ L)C~ ~ o tD O ~
LO~ O . 'O
L(~ ~ o o U~tv) r-~ O
~ CD C~ O ~ o o O
O t~ d~ O O O r-l L~ C~ . O
.,~ ~ O0~ 00 0~ ~
~i O ~ o~ ~o ~C) O O O O
~i CO o(v~lC~ ;I co ~_ ~ o ~
~1 O
~ O ' 0~ ~ 0 O O ~ 0 O ~ C~J ~ I ~ C~ ~1 O ~1 ~ 0~ C~ O
C~ ~ Lt~ CO~I ~ O O ~ 0 . o C~l C`;l ~ ~ . ~ O
O ~ ~ O ~ C~ ~ LO O
. . . ¢ 4-1. ~ C~ . O
E~ O ~o c~o ~ Ir~ Ln ~ o r-l CO 0Lt~ C~ C~ O ,_1 O ~1 ~ 00 C~ . O
0 ~ ~ . o . ~ O ~ o o O O ~ O
~ C`3 ~-1 ~ C~ ~ o 7 O ' O
.~ S~ L~O~ L~O~
:13 ~ ~
~ E~ ~ ~ U ~ ~ ¢ ~ ~ ~ ~ U ¢ ¢ ~ E~ E~
O L~
O
s o ~o ~ o o o o `i c~~ o o o o r-l~ ~ NC~~ ~D O r-l co ~ O
O o o. O O ~ O
~Do C`~ C~ ~ o . O
~C~C~lC~l0~)O ~
~0 ~ O
I~ o oo O O ~ O
Il~ OO~ C~ ~ o . o O
O ,~C~I C~ C`:l ~ O .-~
00 C~ O
O C~ ~ ~ U~ ~ O
~ ~ C~C\I C~l ~ O ~1 O C!:)o~D O .-1 O
~:;~ o ~D ~ o . o O
.~ ~DC~~~:Y O ~
.~ ~ ~ ~ O
O O o ~ ~ O O~ O
t~ N .~ C`J ~ o. . O
~ ~ C~C~l~ ~O
.~ ~C~J O
¢ O ~ O~~D O ~ O
~1. ~ ~ o ~ . O
E~ ~ ~ ~ c~l cr~ ~ O. ~
~D ~ O
o O CO O ~ O
~E~ ~ ~ ~a ~ ¢¢¢~ E~
~ u~ O LO
. MS-1081-CIP ~ ~
~ o o o d1 ~ o, ~ o o o o C'~ . C~ C~l ~ . o o o C~ ~, ~C~ C~l C~ CO o .~
o~ C~ o ' O ~1 ~1 ~ O O O
C~ .o~ C~ ~ . o o Ç~l ~C~ C~ C~lC~ C~ ~
C~l o C~ O
~ O o . O tD O ~ O
. ~ oo C~ CO ~ ~o o ~
o o U~ o ~ o o 'o o C~ Co~l ,1 ~ ~ ~ ~ CO o o ~:1 ~ c~ o E¢~ o o ~ o o o o ~ ~ ~ c~ c~l ~r CD O _~
oo . ~ C~ ~ . . . o c Q ~ ~ Q ~ ¢ ¢ ¢ E~ E~
o ~ .o s ` ' ~, o ~ Y ~ o; C~ o o ~ ~
~o V a~
h ~ O ~ o C`J C~ l O o co ~ ~ o ~1) ~ V OO o) o ~ O ~
h V ~ ~ V O ~
a~ ~ ;~ L~ ~4 CD ~ C`J C~ ' h ~ ~ ~ ~ o oO O o ~
~ O ~ ~ ~ ~ b~
o a~ ~ o o ~ o ~ ~ ~ ~ o C~ o ~ o ~ ~
¢ ~ .~ O C~ O o ~ ~ O C~J CO O
~0~ ~ ~ o . 0 ~ ~ ~ , o ~ CD ~ O
~: ~ co ~ a~ o ~ ~ o ~ o~
o ~ R
t~ h E~ . . . o o o . ~ .+' a~ h . c~l CD CD c~ o CY~ C~ a~ ~
'0 ~ V U~ o~ 0 ~ o ~ C~ c~ o ~:
Q ~ C g U~ ~' g ~ ~ o o ~
~ P ~ C~J c~ ~--1 CO c~) ~ C~ CO ~1 H ~1 .
.t) ~ .
a~ ~ c~ c~ c~ ~ c~ ~ c~l cY~ ~ ~ ~ o~ ~ O c~
~C _ ~ _ _ ~ ~ ~ _ C~ C~
V ~
O ~ O
C~l C~
_ ..... . .. .. .
S
1 Therapeutic utility for the stable solutions o:E
the present invention is shown in the following ex-ample.
E~XAMPLE 25 A solution was prepared having the following composition with contents expressed as weight/volume percent based on total solution volume:
Concen-Ingredient tration _ 10 Tetracycline HCl 1.00 Dioctyl Sodium Sulfosuccinate 2.00 Æthanol 200 Proof 2'1.42 Ascorbic Acid 0.10 Polyethylene Glycol 200 6~0 15 Glyceryl Triacetate 22.10 Diisopropyl Sebacate 44.38 Pight 0.1 ml. portions of the above solution were separately applied to eight 1.4 cm. dia. sections of cadaver leg skin. The solution portions were 20 applied to the surface of the epidermis. As a con-trol, eight 0.1 ml. portions of an aqueous solution of tetracycline hydrochloride available commercially under the trade name "Topicycline" were also sepa-rately applied to eight 1.4 cm. dia. sections of 25 cadaver leg skin. The solution portions were left in contact with the skin for 16 hours and the epidermis was washed with soap and water. The epidermis was then separated from the corium layer of the skin. A
6 mm. dia. punch biopsy sample of each corium layer 30 was then obtained and transferred to a growth plate inoculated with Propionibacterium acnes microorga-nisms. The resulting plates were then incubated for five days under anaerobic conditions at room temper-ature. The resulting organism growth was then , I
_. _ _. . .. . .
1 observed. Any inhibition of growth around -the corium disc will indicate antibiotic therapeutic activity.
The inhibition is expressed as the radius of area having no organism growth. The portions of samples of the solution oE the present invention all had in-hibition areas with radii Erom 9 to 12 mm. None of the prior art samples had any inhibition areas. These results show that the solution of the present in-vention can penetrate through the epidermis and can retain desirable therapeutic acitivity.
The above data on the tetracycline, epitetra-cycline and degradation products (such as anhydro-tetracycline) of the various compositions were ob-tained using a column chromatography-spectrophoto-metric method generally described in the Journal of Pharmaceutical Sciences, Vol. 59, pp. 1480-1482 (1970) or a modification thereof. The modified assay pro-cedure is as follows:
COLUMN CHROMATOGRAPHY SPECTROPHOTOMETRIC PROCEDURE
Several assay solutions are prepared.
1. Butanol-chloroform: mix 100 ml. n-butyl alcohol with 100 ml. chloroform.
2. EDTA: dissolve 37.2 * 0.1 g. (ethylene dinitrilo) tetraacetic acid disodium salt in about 800 ml.
water. Adjust pll to 6.5 + 0.5 with conc. am-monium hydroxide, Dilute to 1 1. with water.
- x 100 = 0% tetracycline hydrochloride at 8 wks. 50 C.
It can thus be seen from the above data that the composition of the present invention is considerably more stable on extended storage than a typical com-parable prior art composition.
Additional examples of varying compositions within the scope of the present invention are listed in the following Table 2. In addition, chemical stability data on a number of these formulations are given in Table 3.
-L~ o ~ O ~ O o O
C~ OtD C~J(:0r-l C~ O r--l ~ CO O
a~ c~ O o~
O o o oCl~ O ~ O
o C~ C~ ~ o ' o O
X ~ L)C~ ~ o tD O ~
LO~ O . 'O
L(~ ~ o o U~tv) r-~ O
~ CD C~ O ~ o o O
O t~ d~ O O O r-l L~ C~ . O
.,~ ~ O0~ 00 0~ ~
~i O ~ o~ ~o ~C) O O O O
~i CO o(v~lC~ ;I co ~_ ~ o ~
~1 O
~ O ' 0~ ~ 0 O O ~ 0 O ~ C~J ~ I ~ C~ ~1 O ~1 ~ 0~ C~ O
C~ ~ Lt~ CO~I ~ O O ~ 0 . o C~l C`;l ~ ~ . ~ O
O ~ ~ O ~ C~ ~ LO O
. . . ¢ 4-1. ~ C~ . O
E~ O ~o c~o ~ Ir~ Ln ~ o r-l CO 0Lt~ C~ C~ O ,_1 O ~1 ~ 00 C~ . O
0 ~ ~ . o . ~ O ~ o o O O ~ O
~ C`3 ~-1 ~ C~ ~ o 7 O ' O
.~ S~ L~O~ L~O~
:13 ~ ~
~ E~ ~ ~ U ~ ~ ¢ ~ ~ ~ ~ U ¢ ¢ ~ E~ E~
O L~
O
s o ~o ~ o o o o `i c~~ o o o o r-l~ ~ NC~~ ~D O r-l co ~ O
O o o. O O ~ O
~Do C`~ C~ ~ o . O
~C~C~lC~l0~)O ~
~0 ~ O
I~ o oo O O ~ O
Il~ OO~ C~ ~ o . o O
O ,~C~I C~ C`:l ~ O .-~
00 C~ O
O C~ ~ ~ U~ ~ O
~ ~ C~C\I C~l ~ O ~1 O C!:)o~D O .-1 O
~:;~ o ~D ~ o . o O
.~ ~DC~~~:Y O ~
.~ ~ ~ ~ O
O O o ~ ~ O O~ O
t~ N .~ C`J ~ o. . O
~ ~ C~C~l~ ~O
.~ ~C~J O
¢ O ~ O~~D O ~ O
~1. ~ ~ o ~ . O
E~ ~ ~ ~ c~l cr~ ~ O. ~
~D ~ O
o O CO O ~ O
~E~ ~ ~ ~a ~ ¢¢¢~ E~
~ u~ O LO
. MS-1081-CIP ~ ~
~ o o o d1 ~ o, ~ o o o o C'~ . C~ C~l ~ . o o o C~ ~, ~C~ C~l C~ CO o .~
o~ C~ o ' O ~1 ~1 ~ O O O
C~ .o~ C~ ~ . o o Ç~l ~C~ C~ C~lC~ C~ ~
C~l o C~ O
~ O o . O tD O ~ O
. ~ oo C~ CO ~ ~o o ~
o o U~ o ~ o o 'o o C~ Co~l ,1 ~ ~ ~ ~ CO o o ~:1 ~ c~ o E¢~ o o ~ o o o o ~ ~ ~ c~ c~l ~r CD O _~
oo . ~ C~ ~ . . . o c Q ~ ~ Q ~ ¢ ¢ ¢ E~ E~
o ~ .o s ` ' ~, o ~ Y ~ o; C~ o o ~ ~
~o V a~
h ~ O ~ o C`J C~ l O o co ~ ~ o ~1) ~ V OO o) o ~ O ~
h V ~ ~ V O ~
a~ ~ ;~ L~ ~4 CD ~ C`J C~ ' h ~ ~ ~ ~ o oO O o ~
~ O ~ ~ ~ ~ b~
o a~ ~ o o ~ o ~ ~ ~ ~ o C~ o ~ o ~ ~
¢ ~ .~ O C~ O o ~ ~ O C~J CO O
~0~ ~ ~ o . 0 ~ ~ ~ , o ~ CD ~ O
~: ~ co ~ a~ o ~ ~ o ~ o~
o ~ R
t~ h E~ . . . o o o . ~ .+' a~ h . c~l CD CD c~ o CY~ C~ a~ ~
'0 ~ V U~ o~ 0 ~ o ~ C~ c~ o ~:
Q ~ C g U~ ~' g ~ ~ o o ~
~ P ~ C~J c~ ~--1 CO c~) ~ C~ CO ~1 H ~1 .
.t) ~ .
a~ ~ c~ c~ c~ ~ c~ ~ c~l cY~ ~ ~ ~ o~ ~ O c~
~C _ ~ _ _ ~ ~ ~ _ C~ C~
V ~
O ~ O
C~l C~
_ ..... . .. .. .
S
1 Therapeutic utility for the stable solutions o:E
the present invention is shown in the following ex-ample.
E~XAMPLE 25 A solution was prepared having the following composition with contents expressed as weight/volume percent based on total solution volume:
Concen-Ingredient tration _ 10 Tetracycline HCl 1.00 Dioctyl Sodium Sulfosuccinate 2.00 Æthanol 200 Proof 2'1.42 Ascorbic Acid 0.10 Polyethylene Glycol 200 6~0 15 Glyceryl Triacetate 22.10 Diisopropyl Sebacate 44.38 Pight 0.1 ml. portions of the above solution were separately applied to eight 1.4 cm. dia. sections of cadaver leg skin. The solution portions were 20 applied to the surface of the epidermis. As a con-trol, eight 0.1 ml. portions of an aqueous solution of tetracycline hydrochloride available commercially under the trade name "Topicycline" were also sepa-rately applied to eight 1.4 cm. dia. sections of 25 cadaver leg skin. The solution portions were left in contact with the skin for 16 hours and the epidermis was washed with soap and water. The epidermis was then separated from the corium layer of the skin. A
6 mm. dia. punch biopsy sample of each corium layer 30 was then obtained and transferred to a growth plate inoculated with Propionibacterium acnes microorga-nisms. The resulting plates were then incubated for five days under anaerobic conditions at room temper-ature. The resulting organism growth was then , I
_. _ _. . .. . .
1 observed. Any inhibition of growth around -the corium disc will indicate antibiotic therapeutic activity.
The inhibition is expressed as the radius of area having no organism growth. The portions of samples of the solution oE the present invention all had in-hibition areas with radii Erom 9 to 12 mm. None of the prior art samples had any inhibition areas. These results show that the solution of the present in-vention can penetrate through the epidermis and can retain desirable therapeutic acitivity.
The above data on the tetracycline, epitetra-cycline and degradation products (such as anhydro-tetracycline) of the various compositions were ob-tained using a column chromatography-spectrophoto-metric method generally described in the Journal of Pharmaceutical Sciences, Vol. 59, pp. 1480-1482 (1970) or a modification thereof. The modified assay pro-cedure is as follows:
COLUMN CHROMATOGRAPHY SPECTROPHOTOMETRIC PROCEDURE
Several assay solutions are prepared.
1. Butanol-chloroform: mix 100 ml. n-butyl alcohol with 100 ml. chloroform.
2. EDTA: dissolve 37.2 * 0.1 g. (ethylene dinitrilo) tetraacetic acid disodium salt in about 800 ml.
water. Adjust pll to 6.5 + 0.5 with conc. am-monium hydroxide, Dilute to 1 1. with water.
3. PEG 400: to 80 ml. glycerin add sufficient poly-ethylene glycol 400 to make 100 ml. Mix well.
4. Buffer: to a 500 ml. volumetric ~lask add 25.0 ml. PEG 400 solution and dilute to volume with EDTA solution.
- l6 -1 5. EtOAc: shake 125 ml. ethyl acetate with 5 ml.
water and about 3 g. (ethylene dinitrilo) tetra-acetic acid disodium salt until saturated.
Filter through Whatman No. 1 filter paper.
6. Alkaline ~leO~I: dilute 5.0 ml. COIIC . ammonium hydroxide to 100 ml. witll anllydrous methyl al-cohol.
7. Tetracycline standard: weigh accurately about 70 mg. tetracycline hydrocllloride and quantita-tively transfer to a 100 ml. volumetric flask.
Add 10 ml. methyl alcohol. After dissolution of the tetracycline, dilute to volume with chloro-form.
The diatomaceous earth to be used as the chromat-ographic column support is prepared by adding about 400 ml. water and 400 ml. hydrochloric acid to about 200 g. Johns-Manville Celite 545 and stirring about 15 min. The solids are then filtered and washed with water until the washings are neutral. The Celite is then mixed Witil about 250 ml. ethyl acetate, and 250 ml. methyl alcohol for 15 min. The solids are then filtered and vacuum dried at 60 C.
The chromatographic column is prepared by mixing 20 ml. buf-fer with 40 g. diatomaceous earth (support) until the support is uniformly coated. A disc of Whatman 541 filter paper is placed at the bottom of a 1.3 x 45 cm. glass column fitted wi-th a Teflon stop-cock at its lower end. Weigh 8 + 0.1 g. support and add to the column in three portions. After each por-tion addition, settle the support by firmly tappingthe column on a padded bench top from a height of
- l6 -1 5. EtOAc: shake 125 ml. ethyl acetate with 5 ml.
water and about 3 g. (ethylene dinitrilo) tetra-acetic acid disodium salt until saturated.
Filter through Whatman No. 1 filter paper.
6. Alkaline ~leO~I: dilute 5.0 ml. COIIC . ammonium hydroxide to 100 ml. witll anllydrous methyl al-cohol.
7. Tetracycline standard: weigh accurately about 70 mg. tetracycline hydrocllloride and quantita-tively transfer to a 100 ml. volumetric flask.
Add 10 ml. methyl alcohol. After dissolution of the tetracycline, dilute to volume with chloro-form.
The diatomaceous earth to be used as the chromat-ographic column support is prepared by adding about 400 ml. water and 400 ml. hydrochloric acid to about 200 g. Johns-Manville Celite 545 and stirring about 15 min. The solids are then filtered and washed with water until the washings are neutral. The Celite is then mixed Witil about 250 ml. ethyl acetate, and 250 ml. methyl alcohol for 15 min. The solids are then filtered and vacuum dried at 60 C.
The chromatographic column is prepared by mixing 20 ml. buf-fer with 40 g. diatomaceous earth (support) until the support is uniformly coated. A disc of Whatman 541 filter paper is placed at the bottom of a 1.3 x 45 cm. glass column fitted wi-th a Teflon stop-cock at its lower end. Weigh 8 + 0.1 g. support and add to the column in three portions. After each por-tion addition, settle the support by firmly tappingthe column on a padded bench top from a height of
5-10 cm. Then lightly tamp the support surface with ~IS-1081-CIP
1 a glass tamping rod having a diameter slightly less than the internal diameter o-f the column. The final support height in the column should be about 10.5-11.5 cm.
A sample solution for assay is prepared by ac-curately transferring a portion of sample equivalent to 15-20 mg. tetracycline (e.g. 9 2.0 ml. for 1% so-lution) to a 25 ml. volumetric flask. Dilute to volume with buf-fer. Transfer exactly 1.0 ml. sample solution to a 50 ml. beaker. Mix thoroughly with 2.0 + 0.1 g. support. Quantitatively transfer the support to the column. Wash the beaker with 0.5 -~0.1 g. support and add to the column. Tamp well.
Add 20 ml. EtOAc solution to the top of the column.
Collect exactly 10 ml. eluate in a 10 ml. volumetric flask. This Cut l will contain the degradation prod-ucts, such as anhydrotetracycline. Change to a 25 ml.
graduated cylinder for the Cut 2 receiver. When the solvent level~ in the column drops to the level of the 20 support~ add 60 ml. chloroform to the column. Collect exactly 20 ml. eluate (Cut 2). Replace the receiver with a 50 ml. volume-tric flask. Collect the remain-ing chloroform eluate (Cut 3) until the chloroform level in the column drops to the level of the support.
25 Replace the 50 ml. volumetric flask with a 10 ml.
graduated cylinder. Add 40 ml. butanol-chloroform to the column and collect 8.0 ml. eluate (Cut 4). Tetra-cycline is in Cuts 2, 3 and 4. Add Cut 4 to Cut 3.
Collect the remaining eluate in a 50 ml. graduated 30 cylinder. This final Cut 5 contains the epitetra-cycline.
The absorbance of Cut 1 at 438 nm in a 1 cm cell is determined versus chloroform. The absorbance of standard tetracycline is obtained by transferring _... ...
1 exactly 2.0 ml. standard tetracycline solution to a 100 ml. volumetric flask, adding about 90 ml. chloro-form and 2.0 ml. alkaline MeOH, mixing and diluting to volume with chloroform. The absorbance of the result-ing solution within 10 min. at 366 nm in a 1 cm. cellversus chloroform is then measured. Cut 2 is mixed well with 2.0 ml. alkaline MeOH and its absorbance is measured in a l cm. cell within lO min. at 366 nm.
versus chloroform. Cut 3 is mixed with 2.0 ml. alka-10 line MeOH and diluted to volume with chloroform. Itsabsorbance is measured within 10 min. at 366 nm. in a 1 cm. cell versus chloroform. Mix Cut 5 with 2.0 ml.
alkaline MeOH and record the final volume. Its ab-sorbance is measured within 10 min. at 366 nm. in a 15 1 cm. cell versus chloroform.
The concentrations of anhydrotetracycline, tetra-cycline and epitetracycline are calculated as follows:
Anhydrotetracycline ~AI`C) Abs. Cut l vol. Cut l sample 0.0185 x sample vol. x dilution x 1000 mg ATC/ml.
mg. ATC/ml./theoretical tetracycline, mg./ml. x 100 =
% ATC
1 Tetracycline (TC~
A) Abs. Cut 2 wt.std.,mg. 2 ml.
Abs. Std.TC x lO0 ml. x lO0 ml. x vol. Cut 2 sample sample vol. x dilU-tion = mg. TC/ml.
B) Abs. Cu-t 3 wt.std.,m~. 2 ml.
Abs. Std.TC lO0 mT. x lO0 -ml. x vol. Cut 3 sample sample vol. x dilution mg. TC/ml.
A + B/theoretical TC, mg./mg. x 100 = % TC
Epitetracycline (ETC) Abs. Cut 5 wt.std.,mg. 2 ml. vol Cut 5 Abs. Std.TC x 100 -~ x 100 ml. x sample vol. x dilution mg. ETC/ml.
mg.ETC/ml./theore-tical TC, mg./ml. x 100 = % ETC
If % ATC + % TC + % ETC is less than 90%, the assay should be repeated to confirm the results.
Subsequently, a different analytical procedùre 20 was employed. This reversed pllase high pressure liquid chromatography procedure is as follows.
REVERSED PHASE HPLC
- Several assay solutions are prepared.
1. Perchloric acid stock solution: dilute 43 ml.
70% perchloric acid to 1 liter with water.
2. Mobile Phase A: dilute 100 ml. perchloric acid stock solution to 1 liter with water.
l 3. Mobile Phase B- dilute 100 ml. perchloric acid stock solution to 1 liter with ~IPLC grade aceto-nitrile .
~. Te-tracycline S-tandard, Stock Solution: Accurately weigh about 12 mg. USP reference tetracycline and quantitati~ely transfer to a 50 ml. volumetric ~lask. Dissolve in and dilute to volume with methanol. Store in a ~reezer.
5. Anhydrotetracycline Standard, Stock Solution: Ac-curately weigh about 5 mg. European Pharmacopia reference standard anhydrotetracycline and quan-titatively transfer to a 100 ml. volumetric flask. Dissolve in and dilute to volume with methanol. Store in a ~ree~er.
15 6. Working Standard: On the day of use mix 1 ml.
each of tetracycline and anhydrotetracycline standard stock solutions.
The equipment employed for the assay is well -known IIPLC apparatus equipped with a 10 microliter loop injector, column oven, gradient generator and 20 computing integrator. The accessory chromatographic column is 4 x 250 mm. containing Lichrosorb RP-8, 10 micron. The initial composition of the liquid applied to the column is 89 vol. percent Mobile Phase A and 11 vol. percent Mobile Phase B. The final composition of 25 the liquid applied to the column is 29 vol. percent Mobile Phase A and 71 vol. percent Mobile Phase B.
The gradient rate is 3 vol. percent/min. and the flow rate is 2 ml./min. The temperature is ~0 C., the de-tection wavelength is 275 nm. and the sensitivity is 30 0.1 AUFS.
~S-10~1-CIP
1 A sample solution for assay is prepared by transferring 1.0 ml. sample to a 25 ml. volumetric flask and diluting to volume with methanol. Filter all samples and standards through Whatman G~/F
filter paper prior to injection. Make duplicate in-jections of thc working standard and single in-jections of each sample. After each injection im-mediately start the gradient and the integrator.
Allow at least 8 min. at initial conditions, for 10 equilibration~ after each gradient run before rnaking the next injection. The approximate retention times are as follows:
Epitetracycline 480 seconds Tetracycline 540 seconds Epianhydrotetracycline 810 seconds Anhydrotetracycline 860 seconds The concentrations of tetracycline, epitetra-cycline, anhydrotetracycline and epianhydrotetra-cycline are calculated as follows:
20 Tetracycline (TC) Peak area sample TC x wt. TC Std.(gm.) x 25 x 100 Peak area STd. TC x 50 ml. x 2 x 1 mi.
% TC (w/v) Epitetracycline (ETC) 25 Peak area sample_ETC x wt. TC std.(gm.) x Peak area std. TC x 50 ml. x 2 x 25 ml. x 100 x 1.293 = % ETC (w/v) 1 Anhydrotetracycline (ATC) Peak area sample ATC x wt. ATC std. (gm.) x Peak area std. ATC x 100 x 25 ml. lx lO0 = % ATC ~w/
Epianhydrotetracycline ~EATC~
Peak area sample EATC x wt. ATC std. ~m.) x Peak area std. ATC x 100 ml. x 25 ml. x 100 x 1-08l = % EATC ~w/Y) Both USP Reference Standard Tetracycline and European Pharmacopia Reference Standard Anhydro-tetracycline may contain small amounts of the corres-ponding epimer. When this occurs, a corrected peak 15 area (P.A.) must be used. This is determined as follows.
Corrected P.A.Std. TC = P.A.Std. TC ~ 1.293 x P.A.ETC, Corrected P.A.Std.ATC = P.A.Std.ATC ~ 1.08~ x P.A.EATC.
While employing the aboye HPLC procedure, it was 20 found that the equilibrium weight ratio of tetra-cycline:epitetracyciine is actually about 3:4. There-fore, whenever this ratio is maintained over extended storage periods, it indicates a stable tetracycline composition.
Additional examples of yarying compositions with-in the scope of the present inYention are listed in the following Table 4. In addition, chemical sta-bility data on several of these formulations are given in Tables 5 and 6. The HPLC assay procedure 30 was used to measure the concentrations of tetra-cycline and epitetracycline shown in Tables S and 6.
.
~ o ~` . ~.o - 2 _ ~ o u~ . a~ ~`1 ~- . . .. o H t~) H t~ ~ ~ 1~ t~ ~ O H
¢ (~1 O
~ ~ O ~ O ~ ~
H --1 e:~ ~1 ~I O t~) ~ O O
~: I~ O t~J 00 .
E- o . . . ~D O~ O
~1 . a~ ~1 ~ . . . o c~ tr) ~t~ ~ (~3 t~ a o 00 ~ O
t ~ ~
O O O ~ ~ t`3 O O O O
E-~1 t~ ~ O
O . ~ O O ~ ~ O
O~ . ~O d . . . . O
~I ~ ~1:7 ~I ~ ~O O ' ~1 00 ~ O
Z; ~ ~
O O . . O O ~ O
~-I 00 . ~D ~ . . . O
Z ~ ~ ~ ~ ~ ~t O ~
' 00 ~1 O
~ Z ~ 'i ~--1 H O . . O O H O
~ I~ . ~e:i' . . . O
¢ :C~ ~ ~ ~ ~ ~ O ~
00 ~ O
~ ~ . ~ . .
O O . . O O ~ O
~ . ~o d- . . . O
V~ ~`I,_( ~ ~`I ~ h ~ o ~1 'C h a) ~ ~ a~ a) ~ . ~
~1 ~ ,~ ~ O
~ ~ rl ~ rl O U~ ~ ~ ~ o .
rT l ~ a~ .9 ~ o ~
u ~ ~ ~ ::~ ~ ~ 5 ~ ,1 ~ a~ ~ o ~1 o , v o O ~ U ~ a~ u ~ rcJ O U~ ~ h O O Cl~ ~ ~ O '~ V~ O O C~
V)r~ h U tl~ ! ~1 a) U U ~1 D ~ J~ t~ ~) O U ~ ~1 ~ :~ t~ C
rl ~ ~ D ~1 ~ O ~ ~1 r-l t~l ~ ~1 ~ ~
~4 ~ X a,~ ~ ~ u $~ a,) ~ ~ ~ ~ o v ~ ~ /
~ ~ ~ u~ ¢ c~ ~ ~ ~ O ~ ~ ~ o ~ E~
¢ ¢ a) ,_,.,~ r-~ ~LI ~ ~I U S~ a> ~-t) .,~
S~ rl ~l ~1 O l ~ ~) U O ~ ~I t~ U ~ ~ E-l a) ~1 ~:1 ~ ~ V~ ~ ¢ O ~ Z a~ o I ¢ P~ ~q z ,-1 ~ ~U v~ ~o4 ~ ) v~ ~( ~Jl U~ o U~ ~P~ O ~ U ~/ ~. ~1 ¢ - ~1 :~ ~ ~ 1 ~ ~ ~ ::) ~1 41 ~ .C ~ U ~d r~ ~ ~ P~
z U~ U U O P~ ~ h ~ O O ~ O :~ O ~ ~ ~ ~ ,D ~ t~ ~Ll O a~ ~ ~ a~ o o ~ ~ c~ ~ ~ ~> ~ ~ a~ a~ ~ ~ ,~ 5~ 1 H ~1 U h :~ u) u~ ; :; cd o ::~ u ~ ::~ ~ t~ ~C O
E-~ ~ ~ ~ 5 ,~ ~,1 ~ O c~ ~ E; ~ O 5 ~1 ~ o U U
H ~3 ~1 ~1) ~ . ,1 . ~1 --1 ~ ~ ~ I ;~ ~ ~1 ~ ~) . ~ ~1 U~ U~ ~ ¢
1~ ~ ~ E-~ ~ ~ 4 ~: ~ Z ~ ~ ~ ~4 ~ J~ ¢ ¢ E-~ ~
~ ~C a~ o o o o , ~: o ¢~ E~ Z Z Z Z . ¢ E~
o n o ~
-~j o ~ a-~ o o~ al ~>~
O r~cr, ~ I
C~ ~. . . . . . .,~
t ~1t~ ~ o . d u~ o h O
t ~ h U~
Z 0 ~0 1~ ~ h a> ~:4 ~ .
~ h ~ G- o ~ ~ t~
~ U~ .O ~ ~ `D O O ~ ~ ~0 ~_ ~~o ~~t ~ .
O. . .~ . .
E~ ~J~1~ `D o ~ n O ( ) h t~
o ~ O ~
1--l h td ,C h IY a) . .,t ~ o 0~ . ~ ~4 U~ o ~`
~a ~ ~ u~ a~^
U~ ~ ~ ~ ~3 ~ ~ ~ oo ~ 11 ~ -O O ~ cn r~ ~t O I~
~ ~C) .. . .o ~ *
~ ~ ~ ~ ~ U~ o ~ ~ o ~: ~o ¢ ~ . td E--Z D O . O
tdu) O~ . .. ~
¢ ~ ~; O a~ ~I td -~; ~ o r~ ~ h ~
E~ ~ ~ ~` ,1 ~ O
1~ 0 o . . .h td ~
~ ~ ~ o . ~ 1 ~ ' td t~ O ~d ::~
h . . ~ h O ~c ~ td h ~1 ~c . . . . .. ~ u~
E~ ~. . ,_/. ,_~ . ~. ,_~ ,~ . ~ ~ ~1 0 Z ~ ~ ~I t~~I t~~1 ~ ~ t~~ t~.--I t~ a~ h ~Ll~ t~ :C O ~7 ~ O t~ I~ o t~ ~C O ~ ~ O t~ I~ o - ~ o ~ 4~
~ ~2: ~ :~ ~ ::~ .~ ~ ,1 ::~ ,1 ::C r~ .,~ ~:4 0 3:~. ~ ~ C_7 ~ C_~ ~ ~ ~ ~.3 ~ . ~ ~ .~ ,Q, O
U~ ~ ~ t~ ~ td t~ ~ td t~ E~ td t~ E~ td t~ td ~ E~ td E- ~ t:~ E--~ ~ E--~ ~ ~_ ~ F~ E~ rll Fr~ E--~ t:t~ O h-~l ¢ E-- td ~d O
U) F :l ~h E~ u~ h :~
O ~d a~ 11 .
~1 n ~D O ~ ,C ,~
O 1~ o L~
s o I
h ~ a~ O O ol )~ ~ O ~i ~rl ~ ,~;
r~
~ ~1 . ~ 10 0 0 U) CO
C ~1 . o a) t~
E~ ~1 ~1 O 1~ h L~ ~ Ci~ ~1 ; 1: td ~ 0 0 1-1 ~rl ~ W) 1=1 . h ~ G O
1~ ~ O 1/1 o .~t ,, a) o~
~1 ~ ~ .~ ~ h ~ ~ O
o o o~ ~ o ~ -1 ~ ~ ~ U) E
O
~ 1 :~ td 4~ h ~ O O ~1 I~ . ~O h O P~ ~ cd ,~
. ~ ~ a~ O U~
Z . . t-d b4 ~ Ui h 1~ ~ ~`J t~ U) td ^
L~ ~ U~ ~ ,CI, a) o c~ O ~ ~ u ~I a~
c~ ~ -¢ (J~ h~1 ~ h ~1 ~ O
~:1 ~ . :~:~ h t~ V :~ ~I C~
¢ E~ .~; oo ~)O P. ~1 ~ p.~ ~I E~
E~ ~ . . ~ ~ ~ o E~ O ~ . ~1 ~ 1 h ~1 ~rl ~ 4~
`D O O ~ O ~ .rl h p~ ~:1 rC O
~ ~ ~ ~ ~ 4~ o u E~
C~7 ~1 5~
¢ ~ o t~ t).O
E- ~ ~ ~ ~ U ~ ~d ~ ~ O ~
F~ . ~ ~ ~d h u) O O ~:
C~ ~ ~3 ~J~ ~1 t~l `.1 ~>
~ ~ a~ ~ , ~ ~ U
t~l ~ . ~ C ~ ~3 h : ~:4 o oo ;~
~1 cs- a~~ v a) a) o h P~
E-- . ~d o ~ ~ u) h :Z . O
~1 . h ~ U U ~,~ O
¢ . ~ ~C~o~ a~o ~ .Y ~ U~l h H 3 1--¦1`~)U) tl~ t~ ~) ~1 a~
~ mh h ~ 0 3: ;C
O' ~ o ~u) ~ ~ a~
.~ ~ ~ o ~
* ~ h~i 0 3 ~ ~ Z 1~
. ~1oO~-l 0 E~ 0 ~1 r-l h Z ~ ~ o c~
H ,D O ~ V~
~ t~ ~ r~0 a~ u a~ ~ ~~ ~ ~I c~ h r~
~1 :~
Pi t~ ~ O h ~ U 0 El H ~ t~ ~I E--~ O O ~ ,~
~d ~4 E~
~C O -1 ~t I~ ~ ~ O ~ t~ ~) ~: O
~1 r-l ~`1 0~ ~`1 1'~) ~ ~) ~r) ~ ~1~ ~c t.) td t) .
o n o o~ 0 ~
o ~o V
CO o~
~ ~ Ln o ~
~ ~ ~t CO o~
~ U~ oo oo oo o~
H
O . ~ ~O ~
~ ~ . . .
~ O O O
e ~ . 5 H ~ O
~ ~ ~ O
H
O ~ ~ ~ ~D O ~ ~ ~ O
~1 ~1 ~ ~`I ~`I ~`1 ~ ~) t~ t~') E-o ~1 ~ -"
s 1 It can thus be seen -from the above data that the compositions of the present invention are considerably more stable on extended storage than a typical prior art composition.
1 a glass tamping rod having a diameter slightly less than the internal diameter o-f the column. The final support height in the column should be about 10.5-11.5 cm.
A sample solution for assay is prepared by ac-curately transferring a portion of sample equivalent to 15-20 mg. tetracycline (e.g. 9 2.0 ml. for 1% so-lution) to a 25 ml. volumetric flask. Dilute to volume with buf-fer. Transfer exactly 1.0 ml. sample solution to a 50 ml. beaker. Mix thoroughly with 2.0 + 0.1 g. support. Quantitatively transfer the support to the column. Wash the beaker with 0.5 -~0.1 g. support and add to the column. Tamp well.
Add 20 ml. EtOAc solution to the top of the column.
Collect exactly 10 ml. eluate in a 10 ml. volumetric flask. This Cut l will contain the degradation prod-ucts, such as anhydrotetracycline. Change to a 25 ml.
graduated cylinder for the Cut 2 receiver. When the solvent level~ in the column drops to the level of the 20 support~ add 60 ml. chloroform to the column. Collect exactly 20 ml. eluate (Cut 2). Replace the receiver with a 50 ml. volume-tric flask. Collect the remain-ing chloroform eluate (Cut 3) until the chloroform level in the column drops to the level of the support.
25 Replace the 50 ml. volumetric flask with a 10 ml.
graduated cylinder. Add 40 ml. butanol-chloroform to the column and collect 8.0 ml. eluate (Cut 4). Tetra-cycline is in Cuts 2, 3 and 4. Add Cut 4 to Cut 3.
Collect the remaining eluate in a 50 ml. graduated 30 cylinder. This final Cut 5 contains the epitetra-cycline.
The absorbance of Cut 1 at 438 nm in a 1 cm cell is determined versus chloroform. The absorbance of standard tetracycline is obtained by transferring _... ...
1 exactly 2.0 ml. standard tetracycline solution to a 100 ml. volumetric flask, adding about 90 ml. chloro-form and 2.0 ml. alkaline MeOH, mixing and diluting to volume with chloroform. The absorbance of the result-ing solution within 10 min. at 366 nm in a 1 cm. cellversus chloroform is then measured. Cut 2 is mixed well with 2.0 ml. alkaline MeOH and its absorbance is measured in a l cm. cell within lO min. at 366 nm.
versus chloroform. Cut 3 is mixed with 2.0 ml. alka-10 line MeOH and diluted to volume with chloroform. Itsabsorbance is measured within 10 min. at 366 nm. in a 1 cm. cell versus chloroform. Mix Cut 5 with 2.0 ml.
alkaline MeOH and record the final volume. Its ab-sorbance is measured within 10 min. at 366 nm. in a 15 1 cm. cell versus chloroform.
The concentrations of anhydrotetracycline, tetra-cycline and epitetracycline are calculated as follows:
Anhydrotetracycline ~AI`C) Abs. Cut l vol. Cut l sample 0.0185 x sample vol. x dilution x 1000 mg ATC/ml.
mg. ATC/ml./theoretical tetracycline, mg./ml. x 100 =
% ATC
1 Tetracycline (TC~
A) Abs. Cut 2 wt.std.,mg. 2 ml.
Abs. Std.TC x lO0 ml. x lO0 ml. x vol. Cut 2 sample sample vol. x dilU-tion = mg. TC/ml.
B) Abs. Cu-t 3 wt.std.,m~. 2 ml.
Abs. Std.TC lO0 mT. x lO0 -ml. x vol. Cut 3 sample sample vol. x dilution mg. TC/ml.
A + B/theoretical TC, mg./mg. x 100 = % TC
Epitetracycline (ETC) Abs. Cut 5 wt.std.,mg. 2 ml. vol Cut 5 Abs. Std.TC x 100 -~ x 100 ml. x sample vol. x dilution mg. ETC/ml.
mg.ETC/ml./theore-tical TC, mg./ml. x 100 = % ETC
If % ATC + % TC + % ETC is less than 90%, the assay should be repeated to confirm the results.
Subsequently, a different analytical procedùre 20 was employed. This reversed pllase high pressure liquid chromatography procedure is as follows.
REVERSED PHASE HPLC
- Several assay solutions are prepared.
1. Perchloric acid stock solution: dilute 43 ml.
70% perchloric acid to 1 liter with water.
2. Mobile Phase A: dilute 100 ml. perchloric acid stock solution to 1 liter with water.
l 3. Mobile Phase B- dilute 100 ml. perchloric acid stock solution to 1 liter with ~IPLC grade aceto-nitrile .
~. Te-tracycline S-tandard, Stock Solution: Accurately weigh about 12 mg. USP reference tetracycline and quantitati~ely transfer to a 50 ml. volumetric ~lask. Dissolve in and dilute to volume with methanol. Store in a ~reezer.
5. Anhydrotetracycline Standard, Stock Solution: Ac-curately weigh about 5 mg. European Pharmacopia reference standard anhydrotetracycline and quan-titatively transfer to a 100 ml. volumetric flask. Dissolve in and dilute to volume with methanol. Store in a ~ree~er.
15 6. Working Standard: On the day of use mix 1 ml.
each of tetracycline and anhydrotetracycline standard stock solutions.
The equipment employed for the assay is well -known IIPLC apparatus equipped with a 10 microliter loop injector, column oven, gradient generator and 20 computing integrator. The accessory chromatographic column is 4 x 250 mm. containing Lichrosorb RP-8, 10 micron. The initial composition of the liquid applied to the column is 89 vol. percent Mobile Phase A and 11 vol. percent Mobile Phase B. The final composition of 25 the liquid applied to the column is 29 vol. percent Mobile Phase A and 71 vol. percent Mobile Phase B.
The gradient rate is 3 vol. percent/min. and the flow rate is 2 ml./min. The temperature is ~0 C., the de-tection wavelength is 275 nm. and the sensitivity is 30 0.1 AUFS.
~S-10~1-CIP
1 A sample solution for assay is prepared by transferring 1.0 ml. sample to a 25 ml. volumetric flask and diluting to volume with methanol. Filter all samples and standards through Whatman G~/F
filter paper prior to injection. Make duplicate in-jections of thc working standard and single in-jections of each sample. After each injection im-mediately start the gradient and the integrator.
Allow at least 8 min. at initial conditions, for 10 equilibration~ after each gradient run before rnaking the next injection. The approximate retention times are as follows:
Epitetracycline 480 seconds Tetracycline 540 seconds Epianhydrotetracycline 810 seconds Anhydrotetracycline 860 seconds The concentrations of tetracycline, epitetra-cycline, anhydrotetracycline and epianhydrotetra-cycline are calculated as follows:
20 Tetracycline (TC) Peak area sample TC x wt. TC Std.(gm.) x 25 x 100 Peak area STd. TC x 50 ml. x 2 x 1 mi.
% TC (w/v) Epitetracycline (ETC) 25 Peak area sample_ETC x wt. TC std.(gm.) x Peak area std. TC x 50 ml. x 2 x 25 ml. x 100 x 1.293 = % ETC (w/v) 1 Anhydrotetracycline (ATC) Peak area sample ATC x wt. ATC std. (gm.) x Peak area std. ATC x 100 x 25 ml. lx lO0 = % ATC ~w/
Epianhydrotetracycline ~EATC~
Peak area sample EATC x wt. ATC std. ~m.) x Peak area std. ATC x 100 ml. x 25 ml. x 100 x 1-08l = % EATC ~w/Y) Both USP Reference Standard Tetracycline and European Pharmacopia Reference Standard Anhydro-tetracycline may contain small amounts of the corres-ponding epimer. When this occurs, a corrected peak 15 area (P.A.) must be used. This is determined as follows.
Corrected P.A.Std. TC = P.A.Std. TC ~ 1.293 x P.A.ETC, Corrected P.A.Std.ATC = P.A.Std.ATC ~ 1.08~ x P.A.EATC.
While employing the aboye HPLC procedure, it was 20 found that the equilibrium weight ratio of tetra-cycline:epitetracyciine is actually about 3:4. There-fore, whenever this ratio is maintained over extended storage periods, it indicates a stable tetracycline composition.
Additional examples of yarying compositions with-in the scope of the present inYention are listed in the following Table 4. In addition, chemical sta-bility data on several of these formulations are given in Tables 5 and 6. The HPLC assay procedure 30 was used to measure the concentrations of tetra-cycline and epitetracycline shown in Tables S and 6.
.
~ o ~` . ~.o - 2 _ ~ o u~ . a~ ~`1 ~- . . .. o H t~) H t~ ~ ~ 1~ t~ ~ O H
¢ (~1 O
~ ~ O ~ O ~ ~
H --1 e:~ ~1 ~I O t~) ~ O O
~: I~ O t~J 00 .
E- o . . . ~D O~ O
~1 . a~ ~1 ~ . . . o c~ tr) ~t~ ~ (~3 t~ a o 00 ~ O
t ~ ~
O O O ~ ~ t`3 O O O O
E-~1 t~ ~ O
O . ~ O O ~ ~ O
O~ . ~O d . . . . O
~I ~ ~1:7 ~I ~ ~O O ' ~1 00 ~ O
Z; ~ ~
O O . . O O ~ O
~-I 00 . ~D ~ . . . O
Z ~ ~ ~ ~ ~ ~t O ~
' 00 ~1 O
~ Z ~ 'i ~--1 H O . . O O H O
~ I~ . ~e:i' . . . O
¢ :C~ ~ ~ ~ ~ ~ O ~
00 ~ O
~ ~ . ~ . .
O O . . O O ~ O
~ . ~o d- . . . O
V~ ~`I,_( ~ ~`I ~ h ~ o ~1 'C h a) ~ ~ a~ a) ~ . ~
~1 ~ ,~ ~ O
~ ~ rl ~ rl O U~ ~ ~ ~ o .
rT l ~ a~ .9 ~ o ~
u ~ ~ ~ ::~ ~ ~ 5 ~ ,1 ~ a~ ~ o ~1 o , v o O ~ U ~ a~ u ~ rcJ O U~ ~ h O O Cl~ ~ ~ O '~ V~ O O C~
V)r~ h U tl~ ! ~1 a) U U ~1 D ~ J~ t~ ~) O U ~ ~1 ~ :~ t~ C
rl ~ ~ D ~1 ~ O ~ ~1 r-l t~l ~ ~1 ~ ~
~4 ~ X a,~ ~ ~ u $~ a,) ~ ~ ~ ~ o v ~ ~ /
~ ~ ~ u~ ¢ c~ ~ ~ ~ O ~ ~ ~ o ~ E~
¢ ¢ a) ,_,.,~ r-~ ~LI ~ ~I U S~ a> ~-t) .,~
S~ rl ~l ~1 O l ~ ~) U O ~ ~I t~ U ~ ~ E-l a) ~1 ~:1 ~ ~ V~ ~ ¢ O ~ Z a~ o I ¢ P~ ~q z ,-1 ~ ~U v~ ~o4 ~ ) v~ ~( ~Jl U~ o U~ ~P~ O ~ U ~/ ~. ~1 ¢ - ~1 :~ ~ ~ 1 ~ ~ ~ ::) ~1 41 ~ .C ~ U ~d r~ ~ ~ P~
z U~ U U O P~ ~ h ~ O O ~ O :~ O ~ ~ ~ ~ ,D ~ t~ ~Ll O a~ ~ ~ a~ o o ~ ~ c~ ~ ~ ~> ~ ~ a~ a~ ~ ~ ,~ 5~ 1 H ~1 U h :~ u) u~ ; :; cd o ::~ u ~ ::~ ~ t~ ~C O
E-~ ~ ~ ~ 5 ,~ ~,1 ~ O c~ ~ E; ~ O 5 ~1 ~ o U U
H ~3 ~1 ~1) ~ . ,1 . ~1 --1 ~ ~ ~ I ;~ ~ ~1 ~ ~) . ~ ~1 U~ U~ ~ ¢
1~ ~ ~ E-~ ~ ~ 4 ~: ~ Z ~ ~ ~ ~4 ~ J~ ¢ ¢ E-~ ~
~ ~C a~ o o o o , ~: o ¢~ E~ Z Z Z Z . ¢ E~
o n o ~
-~j o ~ a-~ o o~ al ~>~
O r~cr, ~ I
C~ ~. . . . . . .,~
t ~1t~ ~ o . d u~ o h O
t ~ h U~
Z 0 ~0 1~ ~ h a> ~:4 ~ .
~ h ~ G- o ~ ~ t~
~ U~ .O ~ ~ `D O O ~ ~ ~0 ~_ ~~o ~~t ~ .
O. . .~ . .
E~ ~J~1~ `D o ~ n O ( ) h t~
o ~ O ~
1--l h td ,C h IY a) . .,t ~ o 0~ . ~ ~4 U~ o ~`
~a ~ ~ u~ a~^
U~ ~ ~ ~ ~3 ~ ~ ~ oo ~ 11 ~ -O O ~ cn r~ ~t O I~
~ ~C) .. . .o ~ *
~ ~ ~ ~ ~ U~ o ~ ~ o ~: ~o ¢ ~ . td E--Z D O . O
tdu) O~ . .. ~
¢ ~ ~; O a~ ~I td -~; ~ o r~ ~ h ~
E~ ~ ~ ~` ,1 ~ O
1~ 0 o . . .h td ~
~ ~ ~ o . ~ 1 ~ ' td t~ O ~d ::~
h . . ~ h O ~c ~ td h ~1 ~c . . . . .. ~ u~
E~ ~. . ,_/. ,_~ . ~. ,_~ ,~ . ~ ~ ~1 0 Z ~ ~ ~I t~~I t~~1 ~ ~ t~~ t~.--I t~ a~ h ~Ll~ t~ :C O ~7 ~ O t~ I~ o t~ ~C O ~ ~ O t~ I~ o - ~ o ~ 4~
~ ~2: ~ :~ ~ ::~ .~ ~ ,1 ::~ ,1 ::C r~ .,~ ~:4 0 3:~. ~ ~ C_7 ~ C_~ ~ ~ ~ ~.3 ~ . ~ ~ .~ ,Q, O
U~ ~ ~ t~ ~ td t~ ~ td t~ E~ td t~ E~ td t~ td ~ E~ td E- ~ t:~ E--~ ~ E--~ ~ ~_ ~ F~ E~ rll Fr~ E--~ t:t~ O h-~l ¢ E-- td ~d O
U) F :l ~h E~ u~ h :~
O ~d a~ 11 .
~1 n ~D O ~ ,C ,~
O 1~ o L~
s o I
h ~ a~ O O ol )~ ~ O ~i ~rl ~ ,~;
r~
~ ~1 . ~ 10 0 0 U) CO
C ~1 . o a) t~
E~ ~1 ~1 O 1~ h L~ ~ Ci~ ~1 ; 1: td ~ 0 0 1-1 ~rl ~ W) 1=1 . h ~ G O
1~ ~ O 1/1 o .~t ,, a) o~
~1 ~ ~ .~ ~ h ~ ~ O
o o o~ ~ o ~ -1 ~ ~ ~ U) E
O
~ 1 :~ td 4~ h ~ O O ~1 I~ . ~O h O P~ ~ cd ,~
. ~ ~ a~ O U~
Z . . t-d b4 ~ Ui h 1~ ~ ~`J t~ U) td ^
L~ ~ U~ ~ ,CI, a) o c~ O ~ ~ u ~I a~
c~ ~ -¢ (J~ h~1 ~ h ~1 ~ O
~:1 ~ . :~:~ h t~ V :~ ~I C~
¢ E~ .~; oo ~)O P. ~1 ~ p.~ ~I E~
E~ ~ . . ~ ~ ~ o E~ O ~ . ~1 ~ 1 h ~1 ~rl ~ 4~
`D O O ~ O ~ .rl h p~ ~:1 rC O
~ ~ ~ ~ ~ 4~ o u E~
C~7 ~1 5~
¢ ~ o t~ t).O
E- ~ ~ ~ ~ U ~ ~d ~ ~ O ~
F~ . ~ ~ ~d h u) O O ~:
C~ ~ ~3 ~J~ ~1 t~l `.1 ~>
~ ~ a~ ~ , ~ ~ U
t~l ~ . ~ C ~ ~3 h : ~:4 o oo ;~
~1 cs- a~~ v a) a) o h P~
E-- . ~d o ~ ~ u) h :Z . O
~1 . h ~ U U ~,~ O
¢ . ~ ~C~o~ a~o ~ .Y ~ U~l h H 3 1--¦1`~)U) tl~ t~ ~) ~1 a~
~ mh h ~ 0 3: ;C
O' ~ o ~u) ~ ~ a~
.~ ~ ~ o ~
* ~ h~i 0 3 ~ ~ Z 1~
. ~1oO~-l 0 E~ 0 ~1 r-l h Z ~ ~ o c~
H ,D O ~ V~
~ t~ ~ r~0 a~ u a~ ~ ~~ ~ ~I c~ h r~
~1 :~
Pi t~ ~ O h ~ U 0 El H ~ t~ ~I E--~ O O ~ ,~
~d ~4 E~
~C O -1 ~t I~ ~ ~ O ~ t~ ~) ~: O
~1 r-l ~`1 0~ ~`1 1'~) ~ ~) ~r) ~ ~1~ ~c t.) td t) .
o n o o~ 0 ~
o ~o V
CO o~
~ ~ Ln o ~
~ ~ ~t CO o~
~ U~ oo oo oo o~
H
O . ~ ~O ~
~ ~ . . .
~ O O O
e ~ . 5 H ~ O
~ ~ ~ O
H
O ~ ~ ~ ~D O ~ ~ ~ O
~1 ~1 ~ ~`I ~`I ~`1 ~ ~) t~ t~') E-o ~1 ~ -"
s 1 It can thus be seen -from the above data that the compositions of the present invention are considerably more stable on extended storage than a typical prior art composition.
Claims (3)
1. A stable nonaqueous solution of a tetracycline antibiotic salt comprising a mixture of 0.25 to 2 per-cent tetracycline antibiotic salt, 41 to 88 percent nonaqueous diluent, 4 to 40 percent nonaqueous solvent, 0.3 to 20 percent nonaqueous nonionic solubilizer, 0 to 6 percent nonaqueous anionic solubilizer and 0 to 0.75 percent antioxidant, said percent values being on a weight/weight basis based on the entire solution weight.
2. A stable solution according to Claim 1 wherein the components are present in the amounts of 0.3 to 1.5 percent tetracycline antibiotic salt, 57 to 84 percent nonaqueous diluent, 4 to 30 percent nonaqueous solvent, 2 to 9 percent nonaqueous nonionic solubilizer, 1 to 6 percent nonaqueous anionic solubilizer and 0.03 to 0.2 percent antioxidant.
3. A stable solution according to Claim 1 wherein the components are present in the amounts of 0.5 to 1.25 percent tetracycline antibiotic salt, 60 to 81 percent nonaqueous diluent, 4 to 25 percent nonaqueous solvent, 3 to 9 percent nonaqueous nonionic solubilizer, 1.5 to 4.5 percent nonaqueous anionic solubilizer and 0.1 to 0.2 percent antioxidant.
Applications Claiming Priority (4)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| US19455680A | 1980-10-06 | 1980-10-06 | |
| US194,556 | 1980-10-06 | ||
| US06/262,475 US4376118A (en) | 1980-10-06 | 1981-05-19 | Stable nonaqueous solution of tetracycline salt |
| US262,475 | 1994-06-20 |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CA1188615A true CA1188615A (en) | 1985-06-11 |
Family
ID=26890152
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CA000384103A Expired CA1188615A (en) | 1980-10-06 | 1981-08-18 | Stable nonaqueous solution of tetracycline salt |
Country Status (7)
| Country | Link |
|---|---|
| US (1) | US4376118A (en) |
| EP (1) | EP0049422B1 (en) |
| AU (1) | AU527976B2 (en) |
| CA (1) | CA1188615A (en) |
| DE (1) | DE3173637D1 (en) |
| ES (1) | ES8306995A1 (en) |
| PH (1) | PH17656A (en) |
Families Citing this family (53)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US4701320A (en) * | 1984-11-29 | 1987-10-20 | Lederle (Japan), Ltd. | Composition stably containing minocycline for treating periodontal diseases |
| US4861580A (en) * | 1985-10-15 | 1989-08-29 | The Liposome Company, Inc. | Composition using salt form of organic acid derivative of alpha-tocopheral |
| US5041278A (en) * | 1985-10-15 | 1991-08-20 | The Liposome Company, Inc. | Alpha tocopherol-based vesicles |
| AU1396588A (en) * | 1987-02-20 | 1988-09-14 | Emory University | Antiinfective compounds and method of use |
| US5811088A (en) * | 1987-02-20 | 1998-09-22 | Emory University | Antiinfective compounds and methods of use |
| US5674911A (en) * | 1987-02-20 | 1997-10-07 | Cytrx Corporation | Antiinfective polyoxypropylene/polyoxyethylene copolymers and methods of use |
| US5122519A (en) * | 1989-06-27 | 1992-06-16 | American Cyanamid Company | Stable, cosmetically acceptable topical gel formulation and method of treatment for acne |
| US6933286B2 (en) | 1991-03-19 | 2005-08-23 | R. Martin Emanuele | Therapeutic delivery compositions and methods of use thereof |
| JP2647556B2 (en) | 1991-03-19 | 1997-08-27 | サイトアーレクス・コーポレーシヨン | Polyoxypropylene / polyoxyethylene copolymers with improved biological activity |
| US7202225B1 (en) | 1993-10-15 | 2007-04-10 | Emanuele R Martin | Therapeutic delivery compositions and methods of use thereof |
| USRE38558E1 (en) | 1991-03-19 | 2004-07-20 | Cytrx Corporation | Polyoxypropylene/polyoxyethylene copolymers with improved biological activity |
| CA2154169A1 (en) * | 1993-01-19 | 1994-08-04 | Pieter Vonk | Cleaning composition |
| US7258869B1 (en) * | 1999-02-08 | 2007-08-21 | Alza Corporation | Stable non-aqueous single phase viscous vehicles and formulations utilizing such vehicle |
| US7919109B2 (en) | 1999-02-08 | 2011-04-05 | Intarcia Therapeutics, Inc. | Stable non-aqueous single phase viscous vehicles and formulations utilizing such vehicles |
| US20020077276A1 (en) * | 1999-04-27 | 2002-06-20 | Fredeking Terry M. | Compositions and methods for treating hemorrhagic virus infections and other disorders |
| US6645508B1 (en) | 1999-06-18 | 2003-11-11 | Jivn-Ren Chen | Stable L-ascorbic acid composition |
| US7731947B2 (en) | 2003-11-17 | 2010-06-08 | Intarcia Therapeutics, Inc. | Composition and dosage form comprising an interferon particle formulation and suspending vehicle |
| US11246913B2 (en) | 2005-02-03 | 2022-02-15 | Intarcia Therapeutics, Inc. | Suspension formulation comprising an insulinotropic peptide |
| WO2006083761A2 (en) | 2005-02-03 | 2006-08-10 | Alza Corporation | Solvent/polymer solutions as suspension vehicles |
| US7919483B2 (en) * | 2005-06-24 | 2011-04-05 | Medicis Pharmaceutical Corporation | Method for the treatment of acne |
| US20080242642A1 (en) * | 2007-04-02 | 2008-10-02 | Medicis Pharmaceutical Corporation | Minocycline oral dosage forms for the treatment of acne |
| US7544373B2 (en) * | 2007-04-02 | 2009-06-09 | Medicis Pharmaceutical Corporation | Minocycline oral dosage forms for the treatment of acne |
| US7541347B2 (en) | 2007-04-02 | 2009-06-02 | Medicis Pharmaceutical Coropration | Minocycline oral dosage forms for the treatment of acne |
| US8722650B1 (en) | 2005-06-24 | 2014-05-13 | Medicis Pharmaceutical Corporation | Extended-release minocycline dosage forms |
| US8252776B2 (en) | 2007-04-02 | 2012-08-28 | Medicis Pharmaceutical Corporation | Minocycline oral dosage forms for the treatment of acne |
| US20080241235A1 (en) * | 2007-04-02 | 2008-10-02 | Medicis Pharmaceutical Corporation | Minocycline oral dosage forms for the treatment of acne |
| CN101453982B (en) | 2006-05-30 | 2011-05-04 | 精达制药公司 | Two-piece, internal-channel osmotic delivery system flow modulator |
| ES2422864T3 (en) | 2006-08-09 | 2013-09-16 | Intarcia Therapeutics, Inc | Osmotic release systems and piston units |
| ES2315123B1 (en) * | 2006-09-25 | 2009-12-30 | Divasa-Farmavic, S.A. | STABLE PHARMACEUTICAL COMPOSITIONS OF TETRACICLINES IN SOLUTION, PROCEDURE FOR OBTAINING AND USES. |
| WO2008097851A1 (en) * | 2007-02-02 | 2008-08-14 | Warner Chilcott Company, Inc. | Tetracycline compositions for topical administration |
| US20080188445A1 (en) * | 2007-02-02 | 2008-08-07 | Warner Chilcott Company Inc. | Tetracycline compositions for topical administration |
| US20080241197A1 (en) * | 2007-04-02 | 2008-10-02 | Medicis Pharmaceutical Corporation | Minocycline dosage forms for the treatment of acne |
| RU2440097C2 (en) | 2007-04-23 | 2012-01-20 | Интарсия Терапьютикс, Инк. | Method of treating insulin-independent diabetes and obesity, osmotic delivery system and method for making it |
| WO2009009135A1 (en) | 2007-07-11 | 2009-01-15 | Dow Corning Corporation | Compositions for delivering a drug |
| MX2007009796A (en) | 2007-08-14 | 2009-02-25 | Cell Therapy And Technology S | Gel containing pirfenidone. |
| EP2240155B1 (en) | 2008-02-13 | 2012-06-06 | Intarcia Therapeutics, Inc | Devices, formulations, and methods for delivery of multiple beneficial agents |
| WO2010017310A1 (en) | 2008-08-06 | 2010-02-11 | Medicis Pharmaceutical Corporation | Method for the treatment of acne and certain dosage forms thereof |
| EP3323423B1 (en) | 2009-09-28 | 2020-06-17 | Intarcia Therapeutics, Inc | Rapid establishment and/or termination of substantial steady-state drug delivery |
| US20120208755A1 (en) | 2011-02-16 | 2012-08-16 | Intarcia Therapeutics, Inc. | Compositions, Devices and Methods of Use Thereof for the Treatment of Cancers |
| US9561241B1 (en) | 2011-06-28 | 2017-02-07 | Medicis Pharmaceutical Corporation | Gastroretentive dosage forms for minocycline |
| MX2011007675A (en) | 2011-07-19 | 2012-07-11 | Cell Therapy And Technology S A De C V | Process for manufacturing a pharmaceutical composition in a sustained-release tablet form containing pirfenidone and the application thereof in the regression of chronic renal failure, breast capsular contracture and liver fibrosis in humans. |
| MX346763B (en) | 2012-03-28 | 2017-03-31 | Cell Therapy And Tech S A De C V | Semi-solid topical composition which contains pirfenidone and modified diallyl disulphide oxide (m-ddo) for eliminating or preventing acne. |
| MX356551B (en) | 2012-08-23 | 2018-06-04 | Grupo Medifarma S A De C V Star | Antiseptic, antiseborrheic, exfoliating composition for getting rid of or preventing acne. |
| WO2015075640A1 (en) | 2013-11-20 | 2015-05-28 | Lupin Limited | Stable pharmaceutical formulation(s) of tetracycline antibiotic |
| US9889085B1 (en) | 2014-09-30 | 2018-02-13 | Intarcia Therapeutics, Inc. | Therapeutic methods for the treatment of diabetes and related conditions for patients with high baseline HbA1c |
| JP6793715B2 (en) | 2015-03-23 | 2020-12-02 | バイオファーミクス・インコーポレイテッドＢｉｏＰｈａｒｍＸ， Ｉｎｃ． | Tetracycline skin pharmaceutical composition |
| EP3302354B1 (en) | 2015-06-03 | 2023-10-04 | i2o Therapeutics, Inc. | Implant placement systems |
| WO2017200943A1 (en) | 2016-05-16 | 2017-11-23 | Intarcia Therapeutics, Inc. | Glucagon-receptor selective polypeptides and methods of use thereof |
| USD860451S1 (en) | 2016-06-02 | 2019-09-17 | Intarcia Therapeutics, Inc. | Implant removal tool |
| USD840030S1 (en) | 2016-06-02 | 2019-02-05 | Intarcia Therapeutics, Inc. | Implant placement guide |
| KR20190104039A (en) | 2017-01-03 | 2019-09-05 | 인타르시아 세라퓨틱스 인코포레이티드 | Methods Including Continuous Administration of GLP-1 Receptor Agonists and Co-administration of Drugs |
| MX366086B (en) | 2017-08-15 | 2019-06-27 | Cell Therapy And Tech S A De C V | A semi-solid topical composition containing an antimicrobial agent and pirfenidone for the treatment of chronic skin damage. |
| CN114428140A (en) * | 2022-01-26 | 2022-05-03 | 坛墨质检科技股份有限公司 | Preparation method of tert-butyl hydroquinone standard solution |
Family Cites Families (8)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US3026248A (en) * | 1959-09-11 | 1962-03-20 | Pfizer & Co C | Thioglycerol and formaldehyde sulfoxylate stabilized tetracycline antibiotics in polyhydric alcohol solvents |
| US3219529A (en) * | 1962-10-04 | 1965-11-23 | American Cyanamid Co | Stable tetracycline solutions |
| US3389174A (en) * | 1962-10-04 | 1968-06-18 | American Cyanamid Co | Stable tetracycline solutions |
| US3140232A (en) * | 1962-12-19 | 1964-07-07 | Pfizer & Co C | Color stabilization of tetracycline compositions with polypropylene glycols |
| US3472931A (en) * | 1969-01-17 | 1969-10-14 | Foster Milburn Co | Percutaneous absorption with lower alkyl amides |
| BE790136R (en) * | 1971-11-29 | 1973-02-15 | Esteve Labor Dr | NEW ESTERS AND HEMI-ESTERS OF DICARBOXYLIC ACIDS AND PROCESS FOR THE PREPARATION OF THESE |
| US4000263A (en) * | 1975-05-02 | 1976-12-28 | Westwood Pharmaceuticals, Inc. | Erythromycin solution |
| JPS5598112A (en) * | 1979-01-17 | 1980-07-25 | Sumitomo Chem Co Ltd | Liquid medicine for external use |
-
1981
- 1981-05-19 US US06/262,475 patent/US4376118A/en not_active Expired - Lifetime
- 1981-08-18 CA CA000384103A patent/CA1188615A/en not_active Expired
- 1981-08-18 PH PH26058A patent/PH17656A/en unknown
- 1981-09-23 EP EP81107568A patent/EP0049422B1/en not_active Expired
- 1981-09-23 DE DE8181107568T patent/DE3173637D1/en not_active Expired
- 1981-10-05 AU AU76032/81A patent/AU527976B2/en not_active Ceased
- 1981-10-05 ES ES506009A patent/ES8306995A1/en not_active Expired
Also Published As
| Publication number | Publication date |
|---|---|
| ES506009A0 (en) | 1983-06-16 |
| PH17656A (en) | 1984-10-23 |
| AU527976B2 (en) | 1983-03-31 |
| AU7603281A (en) | 1982-07-15 |
| EP0049422A3 (en) | 1983-01-26 |
| ES8306995A1 (en) | 1983-06-16 |
| US4376118A (en) | 1983-03-08 |
| EP0049422B1 (en) | 1986-01-29 |
| DE3173637D1 (en) | 1986-03-13 |
| EP0049422A2 (en) | 1982-04-14 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CA1188615A (en) | Stable nonaqueous solution of tetracycline salt | |
| US5149719A (en) | Composition for transdermal penetration of medicaments | |
| Goldberg-Cettina et al. | Enhanced transdermal delivery of estradiol in vitro using binary vehicles of isopropyl myristate and short-chain alkanols | |
| SK284622B6 (en) | Pharmaceutical composition comprising anti-oxidant and use of antioxidant to stabilise this composition | |
| DE60312049T2 (en) | STABILIZED PHARMACEUTICAL COMPOSITION CONTAINING AN AMORPHANT AGENT | |
| US5889051A (en) | Stabilization of prostaglandin drug | |
| Kreuter et al. | Distribution and elimination of poly (methyl methacrylate) nanoparticles after subcutaneous administration to rats | |
| IE782130L (en) | Imidazolidinones | |
| AU5909498A (en) | Formulations for amylin agonist peptides | |
| KR940005278A (en) | Pharmaceutical composition | |
| CA1309025C (en) | Topical preparation containing ofloxacin | |
| AU669122B2 (en) | New pyrazine derivatives, their preparation and their use | |
| Cho et al. | Metronidazole phosphate—A water‐soluble prodrug for parenteral solutions of metronidazole | |
| Chen et al. | Determination of fluorescein and fluorescein monoglucuronide excreted in urine | |
| DE3866408D1 (en) | SELF-ADHESIVE DEVICE FOR TRANSDERMAL ADMINISTRATION OF AN ACTIVE SUBSTANCE. | |
| TWI255720B (en) | Novel preparation and administration form comprising an acid-labile active compound and process for production thereof | |
| KR920700647A (en) | Composition for Transdermal Administration | |
| AU633207B2 (en) | Compositions comprising cytotoxic agent and permeation enhancers | |
| US5650149A (en) | Cosmetic composition for proliferating indigenous bacteria on skin | |
| El-Nakeeb et al. | Uptake of griseofulvin by the sensitive dermatophyte, Microsporum gypseum | |
| Baron et al. | Bioactivity of imipenem as a function of medium, time, and temperature | |
| Ponder | THE MECHANISM OF THE INHIBITION OF HEMOLYSIS: III. INHIBITION BY SOLS OF SUBSTANCES RELATED TO CHOLESTEROL | |
| JPS55149209A (en) | Composition for rectal administration | |
| US3651218A (en) | Antifungal compositions | |
| Gupta | Quantitation of glipizide and glyburide in tablets using high performance liquid chromatography |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| MKEX | Expiry | ||
| MKEX | Expiry |
Effective date: 20020611 |