Injectable ready-to-use solutions containing an antitumor anthracycline glycoside

INJECTABLE READY-TO-USE SOLUTIONS CONTAINING AN ANTITUMOR ANTHRACYCLINE GLYCOSIDE

The present invention relates to a storage stable, injectable ready-to-use solution of an antitumor anthracycline glycoside, e.g. doxorubicin, and to a process for preparing such a solution.

The anthracycline glycoside compounds are a well known class of compounds in the antineoplastic group of agents, of which doxorubicin is a typical, and the most widely used, representative: Doxorubicin. Anticancer Antibiotics, Federico Arcamone, 1981, Publ: Academic Press, New York, N.Y.; Adriamycin Review, EROTC International Symposium, Brussels, May, 1974, edited by M. staquet, Publ. Eur. Press Medikon, Ghent, Belg.; and Results of Adriamycin Therapy, Adriamycin Symposium at Frankfurt/Main 1974 edited by M.Ghione, J.Fetzer and H.Maier, publ.: Springer, New York, N.Y.

At present, anthracycline glycoside antitumor drugs, in particular, e.g. doxorubicin, are solely available in the form of lyophilized preparations, which need to be reconstituted before administration. Both the manufacture and the reconstitution of such preparations expose the involved personnel (workers, pharmacists, medical personnel, nurses) to risks of contamination which are particularly serious due to the toxicity of the antitumor substances.

The Martindale Extra Pharmacopoeia 28th edition, page 175 left column, reports on adverse effects of antineoplastic drugs and recommends that ״They must be handled with great care and contact with skin and eyes avoided; they should not be inhaled, care must be taken to avoid extravasation since pain and tissue damage may ensue. Similarly, Scand. J. Work Environ Health vol.10 (2), pages 711984) 74־), as well as articles in Chemistry Industry, Issue July 4, 1983, page 488, and Drug-TopicsMedical-Economics-Co, Issue February 7, 1983, page 99 .64703-3?

report about severe adverse effects observed in medical personnel exposed to use of cytostatic agents, including doxorubicin.

To administer a lyophilized preparation, double handling of the drug is required. The lyophilized cake first has to be reconstituted and then administered. Moreover, in some cases, the complete dissolution of the powder may require prolonged shaking because of solubilization problems. Reconstitution of a lyophilized cake or powder can result in formation of aerosol droplets which can be inhaled or can come into contact with skin or mucous membranes of those handling the solution.

As the risks connected with the manufacture and the reconstitution of a lyophilized preparate would be highly reduced if a ready-to-use solution of the drug were available, we have developed a stable, therapeutically acceptable intravenously injectable solution of an anthracycline glycoside drug, e.g. doxorubicin, whose preparation and administration does not require either lyophilization or reconstitution.

CA 103 183460η reports an investigation into the stability of doxorubicin and seven other anthracyclines. Solutions whose pH had been adjusted with perchloric acid, which is not a physiologically acceptable acid because it precipitates blood proteins, were studied. The anthracycline concentration was 5Mg/ml which is far too low for pharmaceutical use. CA 104 155834q reports an

84703-/ investigation into the chemical stability of six anthracyclines in four infusion fluids whose pH had not been adjusted by an acid or buffer.

Application No. 79129 describes and claims an injectable, ready-to-use, sterile, pyrogen-free, anthracycline glycoside solution which consists essentially of a physiologically acceptable salt of an anthracycline glycoside such as doxorubicin dissolved in a physiologically acceptable aqueous solvent therefor at an anthracycline glycoside concentration of from 0.1 to 50 mg/ml, which has not been reconstituted from a lyophilizate and the pH of which has been adjusted to from 2.5 to 5.0 solely with a physiologically acceptable acid. An especially preferred pH is about 3. The Examples illustrate solutions with pH^,s ranging from 2.62 to 3.14. There is no mention of stabilising agents. Dextrose, lactose and mannitol are mentioned as tonicity adjustment agents but no indication is given of the proportions in which they may be used.

According to the present invention, there is provided a ready-to-use, storage stable, sterile, pyrogen-free, injectable anthracycline glycoside solution which consists essentially of a physiologically acceptable salt of an anthracycline glycoside dissolved in a physiologically acceptable aqueous solvent therefor at an anthracycline glycoside concentration of from 0.1 to 50 mg/ml, which has not been reconstituted from a lyophilizate and the pH of which has been adjusted to from 2.5 to 3.5 by means of a glycine buffer.

It is thus possible to provide solutions which are

׳״ 3=703 84 storage stable and have a commercially meaningful shelflife.

Preferably the solution of the invention is provided in a sealed container, especially one made of glass. The solution can be provided in this way either in a unit dosage form or in a multiple dosage form.

Preferably the anthracycline glycoside is chosen from doxorubicin, 4'-epi-doxorubicin (i.e. epirubicin), 4׳-desoxy-4'-iodo-doxorubicin, daunorubicin and 4-demethoxy-daunorubicin (i.e. idarubicin). Particularly preferred anthracycline glycosides are doxorubicin and 4'-epi-doxorubicin, especially doxorubicin.

Any physiologically acceptable salt of the anthracycline glycoside may be used for preparing the solution of the invention. Examples of suitable salts may be, for instance, the salts with mineral inorganic acids such as hydrochloric, hydrobromic, sulfuric or phosphoric acid and the salts with certain acids such as succinic, tartaric, ascorbic, citric, methanesulfonic or ethanesulfonic acid. The salt with hydrochloric acid is a particularly preferred salt, especially when the anthracycline glycoside is doxorubicin.

Any aqueous solvent which is physiologically acceptable and which is able to dissolve the anthracycline glycoside salt may be used. The solution of the invention may also contain one or more formulation adjuvants such as a co-solubilizing agent (which may be the same as a solvent),

״3־84703 a tonicity adjustment agent, a preservative and a pharmaceutically acceptable chelating agent.

Suitable solvents and co-solubilizing agents may be, for instance, water e.g. Water for Injections; a 0.9% sodium chloride solution, i.e. physiological saline; and aqueous 5% dextrose solution; and mixtures of water and one or more of:

- an aliphatic amide, e.g. Ν,Ν-dimethylacetamide or N-hydroxy-2-ethyl-lactamide;

- an alcohol, e.g. ethanol or benzyl alcohol;

- a glycol or polyalcohol, e.g. propyleneglycol or glycerin;

- an ester of a polyalcohol, e.g. diacetine or triacetine;

- a polyglycol or polyether, e.g. polyethyleneglycol 400 or a propyleneglycol methylether;

- a dioxolane, e.g. isopropylidenglycerin;

- dimethylisosorbide; and

- a pyrrolidone derivative, e.g. 2-pyrrolidone, N-methyl-2pyrrolidone or polyvinylpyrrolidone.

Examples of preferred solvents are water, physiological saline, an aqueous 5% dextrose solution, and mixtures of water and one or more of ethanol, polyethyleneglycol and dimethylacetamide. Water, physiological saline and a 5% dextrose solution are particularly preferred.

Suitable tonicity adjustment agents may be, for instance, physiologically acceptable inorganic chlorides,

e.g. sodium chloride, dextrose, lactose, mannitol, sorbitol

י* 703-3 84

- סand the like .

Preservatives suitable for physiological administration may be, for instance, esters of para-hydroxybenzoic acid (e.g., methyl, ethyl, propyl and butyl esters, or mixtures of them), chlorocresol and the like.

A suitable pharmaceutically acceptable chelating agent may be ethylenediaminotetraacetic acid (EDTA). The chelating agent is included in a minor amount, typically from 0.001 to 0.05% by weight.

The above mentioned solvents, tonicity adjustment agents, preservatives and chelating agents can be used alone or as a mixture of two or more of them.

To adjust the pH within the range of from 2.5 to 3.5 a glycine buffer is added as desired. The range of pH for the ready-to-use solutions of the invention is from 2.5, e.g. from about 2.6, to about about 3.5. A more preferred pH range is from 3 to 3.5. A pH of about 3 is particularly preferred especially where the solution of the invention contains sorbitol, dextrose, lactose or mannitol. Other preferred pH ranges are from greater than 3.14, e.g. from about 3.2, to 3.5. A useful solution with a pH of from 2.62 to 3.14 further comprises a pharmaceutically acceptable chelating agent.

In the solutions of the invention the concentration of the anthracycline glycoside may vary within broad ranges, preferably from 1 mg/ml to 20 mg/ml. The preferred ranges of concentration may be slightly different for different anthracycline glycosides. Thus, for example, preferred concentrations for doxorubicin are from about 2 mg/ml to about 50 mg/ml, preferably from 2 mg/ml to 20 mg/ml, particularly appropriate values being 2 mg/ml and 5 mg/ml. Similar concentrations are preferred also for 4^,-epi-doxorubicin and 4^,-desoxy-4^,-iodo-doxorubicin. Preferred ranges of concentration for daunorubicin and 4-demethoxy-daunorubicin are from 1 mg/ml to 20 mg/ml, concentrations of 1 mg/ml and 5 mg/ml being particularly appropriate.

Suitable packaging for the anthracycline glycoside solutions may be all approved containers intended for parenteral use, such as plastic and glass containers, ready-touse syringes and the like. Preferably the container is a sealed glass container; e.g. a vial or an ampoule. A hermetically sealed glass vial is particularly preferred.

The invention also provides a process for producing a ready-to-use, storage-stable, sterile, pyrogen-free, injectable anthracycline glycoside solution which process comprises dissolving a physiologically acceptable salt of an anthracycline glycoside, which salt is not in the form of a lyophilizate, in a physiologically acceptable aqueous solvent therefor at an anthracycline glycoside concentration of from 0.1 to 50 mg/ml and adding a glycine buffer to adjust the pH of the solution to from 2.5 to 3.5 as desired, the process being effected in such a manner that the resultant solution is sterile and pyrogen-free.

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A suitable process comprises (i) dissolving the physiologically acceptable salt in the physiologically acceptable solvent;

(ii) adding the one or more formulation adjuvants selected from co-solubilizing agents, tonicity adjustment agents, preservatives and pharmaceutically acceptable chelating agents; and (iii) adding the glycine buffer.

Any suitable procedure may be adopted to ensure that the resultant solution is sterile and pyrogen-free. Preferably, the solution is passed through a sterilising filter after addition of the buffer although of course one or more of the materials used may be sterile and pyrogenfree anyway. Where all materials employed are sterile and pyrogen-free, there may then be no need for passing the resultant solution through a sterilising filter.

With the solutions of the invention it is possible to obtain compositions having a very high concentration of the anthracycline glycoside active substance even at 50 mg/ml. This constitutes a great advantage over the presently available lyophilized preparates wherein high concentrations of anthracycline

2־84703 glycoside can only be obtained with difficulty because of solubilization problems encountered in reconstitution, mainly with saline. The presence of the excipient, e.g. lactose, in the lyophilized cake, and its generally high proportion in respect of the active substance, even up to 5 parts of excipient per part of active substance, has a negative effect on solubilization so that difficulties may arise in obtaining dissolution of the lyophilized cake, especially for concentrations of anthracycline glycoside higher than 2 mg/ml.

The solutions of the invention are characterized by a good stability. Solutions in various solvents and with different pH's and concentrations have been found to be stable for long periods at temperatures accepted for the storage of pharmaceutical preparations. This is illustrated in the Examples which follow.

Owing to the well known anti-tumor activity of the anthracycline glycoside active drug substance, the pharmaceutical compositions of the invention are useful for treating tumors in both human and animal hosts. Examples of tumors that can be treated are, for instance, sarcomas, including osteogenic and soft tissue sarcomas, carcinomas, e.g., breast-, lung-, bladder-, thyroid-, prostate- and ovarian carcinoma, lymphomas, including Hodgkin and non-Hodgkin lymphomas, neuroblastoma, melanoma, myeloma, Wilms tumor, and leukemias, including acute lymphoblastic leukemia and acute myeloblastic leukemia.

'״ 84703-3

נ ך ׳־' ן כ

Examples of specific tumours that can be treated are Moloney Sarcona Virus, Sarcoma 180 Ascites, solid Sarcoma 180, gross transplantable leukemia, L 1210 leukemia and lymphocytic P 3BB leukemia.

Inhibition of the growth of a tumour, in particular one of those indicated above, can be achieved by administering to a host suffering from a said tumour an injectable solution according to the invention containing the active drug substance in an amount sufficient to inhibit the growth of said tumour.

The injectable solutions of the invention are administered by rapid e.g. intravenous injection or infusion according to a variety of possible dose schedules.

A Suitable dose schedule for doxorubicin may be, for example, of 60 to 75 mg of active drug substance per m²of body surface given as a single rapid infusion and repeated at 21 days. An alternative schedule may be of 30 mg/m² day be intravenous route .for 3 days, every 28 days. Suitable dosages for 4׳-epi-doxorubicin may be, for instance, of 75 to 90 mg/m² given in a single infusion to be repeated at 21 days, and similar dosages may be useful also for 4׳-desoxy4 ׳-iodo-doxorubicin.

Idarubicin, i.e. 4-demethoxy-daunorubicin, may be, e.g. administered intravenously at a single dose of 13-15 mg/m² every 21 days in the treatment of solid tumours, while in the treatment of leukemias a preferred

3־84703 dose schedule is, e.g., of 10-12 mg/m² day by intravenous route for 3 days, to be repeated every 15-21 days. Similar dosages may be, e.g., followed for daunorubicin.

The following Examples illustrate the invention. Example 1: Doxorubicin.HC1 solutions in sterile water, 5% dextrose or 0.9% saline

Doxorubicin.HC1 was dissolved at a concentration of 2 mg/ml in 1=0.05, pH 2.5 and 3.0 glycine buffers.

Each solution was filtered through a 0.22 gm microporous membrane under nitrogen pressure. 5.0 ml of each solution were stored at 55°C in glass vials of glass type I, 8ml top capacity vial, Teflon (Trade Mark)-faced chlorobutyl rubber bung, aluminium seal. Each solution was analysed at predetermined times (up to 120 hours) for doxorubicin.HC1 assay and pH. The results are shown in Tables 1, 2 and 3 which give the doxorubicin.HC1 residual concentration and percent stability at 55°C, at different pHs and times of storage for sterile water, 5% dextrose and 0.9% saline solutions, respectively.

The doxorubicin.HC1 assays are the mean of three independent determinations performed in accordance with the US Pharmacopoeia (USP high performance liquid chromatography (HPLC) method (USP XXI)). At each pH value, the pseudo-first order rate constants (K_obs) for the degradation were calculated by linear regression analysis of the natural logarithm of the residual concentration of doxorubicin.HC1 ([Dx]_t) versus time as depicted by the following equation:

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In [Dx]^ ⁼ in [Dx]q - Κ_ο^₃ . t

Tables 4, 5 and 6 give the observed rate constants (K_obs) f°^r degradation kinetics of doxorubicin.HC1 at 55*0 and at different pHs for sterile water, 5% dextrose and

0.9% saline solutions, respectively.

water at various pHs

Buffers Tests

Table 1 ־ Accelerated (55‘C) stability data of 2 mg/ml doxorubicin.HCl solutions in sterile

<td> Time (hours)</td>

<td> 0 8 16 24 48 72 120</td>

Doxorubicin.HCl assay

<td> pH 2.5</td><td> . mg/ml</td><td> 1.992</td><td> 1.926</td><td> 1.835</td><td> 1.718</td><td> 1.557</td><td> 1.00</td><td></td>

<td> glycine.HC1</td><td> . % stability</td><td> 100.0</td><td> 96.7</td><td> 92.1</td><td> 86.2</td><td> 78.2</td><td> 50.2</td><td></td>

<td></td><td> PH</td><td> 2.51</td><td> 2.50</td><td> 2.50</td><td> 2.52</td><td> 2.51</td><td> 2.52</td><td></td>

<td></td><td> Doxorubicin.HCl assay</td><td></td><td></td><td></td><td></td><td></td><td></td><td></td>

<td> pH 3.0</td><td> . mg/ml</td><td> 2.003</td><td> 1.958</td><td> 1.881</td><td> 1.831</td><td> 1.696</td><td> 1.525</td><td> 1.258</td>

<td> glycine.HC1</td><td> . 1 stability</td><td> 100.0</td><td> 97.8</td><td> 93.9</td><td> 91.4</td><td> 84.7</td><td> 76.1</td><td> 62.8</td>

<td></td><td> pH</td><td> 3.00</td><td> 3.03</td><td> 3.02</td><td> 3.02</td><td> 3.01</td><td> 3.02</td><td> 3.00</td>

Table 2 Accelerated (55‘C) stability data of 2 mg/ml doxorubicin.HCl solutions in 51 dextrose at various pHs.

Time (hours)

Buffers Tests 0 8 1¢ 24 14___48----72—26—120_

Doxorubicin.HCl assay

<td> pH 2.5</td><td> . mg/ml</td><td> 1.967</td><td> 1.897</td><td> 1.822</td><td> 1.760 1.682</td><td> 1.499 1.305</td>

<td> glycine.HC1</td><td> . ί stability</td><td> 100.0</td><td> 96.4</td><td> 92.6</td><td> 89.5 85.5</td><td> 76.2 66.3</td>

<td></td><td> pH</td><td> 2.56</td><td> 2.56</td><td> 2.56</td><td> 2.58 2.60</td><td> 2.56 2.61</td>

Doxorubicin.HCl assay

<td> pH 3.0</td><td> . mg/ml</td><td> 1.975</td><td> 1.908</td><td> 1.832</td><td> 1.645 1.508 1.344 1.206</td>

<td> glycine.HC1</td><td> . ί stability</td><td> 100.0</td><td> 96.6</td><td> 92.7</td><td> 83.3 76.4 68.0 61.1</td>

<td></td><td> pH</td><td> 3.04</td><td> 3.05</td><td> 3.05</td><td> 3.06 3.00 3.13 3.10</td>

O w I u

<td> Table 3 -</td><td colspan="6"> Accelerated (55’C) stability data of 2 ng/ul doxorubicin.HCl solutions in 0.91 saline</td><td></td>

<td></td><td> at various pHs. •</td><td></td><td> 0</td><td></td><td></td><td></td><td></td>

<td></td><td></td><td></td><td></td><td colspan="2"> Time (hours)</td><td></td><td></td>

<td rowspan="2"> Buffers</td><td rowspan="2"> Tests Doxorubicin.HCl assay</td><td> 0____</td><td> 4 8</td><td> 16</td><td> 24____</td><td> 34 48 72 96 120</td><td></td>

<td></td><td></td><td></td><td></td><td></td><td></td>

<td> pH 2.5</td><td> . mg/ml</td><td> 1.946</td><td> 1.875</td><td> 1.670</td><td> 1.602</td><td> 1.368 1.132</td><td> I</td>

<td> glycine.HCl</td><td> . 1 stability</td><td> 100.0</td><td> 96.3</td><td> 85.5</td><td> 82.3</td><td> 70.3 58.1</td><td rowspan="2"> 1 u in 1</td>

<td></td><td> PH</td><td> 2.59</td><td> 2.59</td><td> 2.59</td><td> 2.58</td><td> 2.62 2.62</td>

<td></td><td> Doxorubicin.HCl assay</td><td></td><td></td><td></td><td></td><td></td><td></td>

<td> pH 3.0</td><td> . mg/ml</td><td> 1.994</td><td></td><td> 1.818</td><td> 1.771</td><td> 1.571 1.375 1.205 1.003</td><td></td>

<td> glycine.HCl</td><td> . 1 stability</td><td> 100.0</td><td></td><td> 91.2</td><td> 88.8</td><td> 78.8 69.0 60.4 50.3</td><td></td>

<td></td><td> pH</td><td> 3.06</td><td></td><td> 3.07</td><td> 3.07</td><td> 3.08 3.13 3.14 3.12</td><td> 00</td>

<td></td><td></td><td></td><td></td><td></td><td></td><td></td><td> *J 0 co</td>

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Table Λ - K values (1/days) for the degradation of doxorubicin.HCI obs

<td> 2</td><td> mg/ml solutions</td><td> in sterile water</td><td> at various pHs at 55^eC</td>

<td> Buffer</td><td> pH_</td><td> 3 K . x 10</td><td> 95% confidence limits</td>

<td></td><td></td><td> ־—obs</td><td></td>

<td> . Glycine-HCl</td><td> 2.5</td><td> 138.3</td><td>.6</td>

<td> (0.05 ש I)</td><td></td><td></td><td></td>

<td> . Glycine-HCl</td><td> 3.0</td><td> 93.1</td><td>.6</td>

(0.05 ־ I) י 703-2 84

Table 5 - values (1/days) for the degradation of doxorubicin.HC1 mg/ml solutions in 5% dextrose at various pHs at 55°C

<td> g-^uffer PH K x 10³ —obs-----</td><td> 95% confidence limits</td>

<td> . Glycine-HCl (I = 0.05)</td><td> 2.5</td><td> 138.7</td><td> 4 - 9.9</td>

<td> • Glycine-HCl (I - 0.05)</td><td> 3.0</td><td> 100.5</td><td> 4 • 5.9</td>

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Table 6 - values (l/days) for the degradation of doxorubicin.HC1 mg/ml solutions in 0.9% saline at various pHs at 55°C

Buffer

95% confidence limis . Glycine-HCl 2.5 (I = 0.05) . Glycine-HCl 3.0 (I = 0.05)

276.5

133.2 .2 .0

Example 2: 4¹-epi-doxorubicin (i.e, epirubicin) solutions

Solutions of epirubicin were prepared in the same fashion as the corresponding doxorubicin solutions of Example 1. They were then tested for stability in the same way. The results are presented in Tables 7 to 12.

Table 7 - Accelerated (55'C) stability data of 2 mg/ml epirubicin.HCl solutions in sterile water at various pHs.

»

Tiiiie (hours)

<td> Buffers</td><td> Tests</td><td> 0_____</td><td> 16</td><td> 24</td><td> 48</td><td> 72</td><td> 96</td><td> JL20</td>

<td> pH 2.5</td><td> Epirubicin.HCl assay . mg/ml</td><td> 2.023</td><td> 1.972</td><td> 1.923</td><td> 1.823</td><td> 1.777</td><td> 1.622</td><td> 1.572</td>

<td> glycine.HCl</td><td> . % initial</td><td> 100.0</td><td> 99.9</td><td> 94.6</td><td> 89.7</td><td> 87.4</td><td> 79.8</td><td> 77.3 _t</td>

<td></td><td> pH</td><td> 2.5</td><td> 2.4</td><td> 2.5</td><td> 2.5</td><td> 2.4</td><td> 2.4</td><td> 2.4 J 1</td>

<td> pH 3.0</td><td> Epirubicin.HCl assay . mg/ml</td><td> 2.055</td><td> 2.008</td><td> 1.917</td><td> 1.764</td><td> 1.709</td><td> 1.571</td><td> 1.494</td>

<td> glycine.HCl</td><td> . $ initial</td><td> 100.0</td><td> 97.7</td><td> 93.3</td><td> 85.8</td><td> 83.2</td><td> 76.4</td><td> 72.7</td>

<td></td><td> pH</td><td> 2.9</td><td> 2.9</td><td> 2.9</td><td> 2.8</td><td> 2.9</td><td> 2.9</td><td> 2.9</td>

!μ ό w I w

Table 8 - Accelerated (55‘C) stability data of 2 mg/ml epirubicin.HCl solutions in 0.9% Sodium Chloride Injection at various pHs.

Time (hours)

<td> Buffers</td><td> Tests 0___</td><td> 4 8</td><td> 16</td><td> 24</td><td> 34</td><td> 48</td><td> 72</td><td> 96</td><td> 120</td><td> 144</td>

<td> 1</td><td> Epirubicin.HCl assay</td><td></td><td></td><td></td><td></td><td></td><td></td><td></td><td></td><td> •</td>

<td> pH 2.5</td><td> . mg/ml 2.077</td><td> 2.066</td><td> 1.987</td><td></td><td></td><td> 1.781</td><td> 1.665</td><td> 1.449</td><td> 1.285</td><td> 1.175</td>

<td> glycine.HC1</td><td> . 1 initial 100.0</td><td> n.d. 99.5</td><td> 95.7</td><td> n.d.</td><td> n.d.</td><td> 85.7</td><td> 80.2</td><td> 69.8</td><td> 61.9</td><td> 56.6 1</td>

<td></td><td> pH 2.4</td><td> 2.4</td><td> 2.4</td><td></td><td></td><td> 2.4</td><td> 2.4</td><td> 2.4</td><td> 2.4</td><td> 2.4 n ס</td>

Epirubicin.HCl assay

<td> pH 3.0</td><td> . mg/ml</td><td> 2.058</td><td> 1.951</td><td> 1.934</td><td> 1.869</td><td> 1.784</td><td> 1.668</td><td> 1.483</td><td> 1.349</td><td> 1.253</td>

<td> glycine.HC1</td><td> . 1 initial</td><td> 100.0</td><td> n.d. n.d. 94.8</td><td> 94.0</td><td> 90.8</td><td> 86.7</td><td> 81.0</td><td> 72.1</td><td> 65.5</td><td> 60.9</td>

<td></td><td> PH</td><td> 3.0</td><td> 2.9</td><td> 2.9</td><td> 2.9</td><td> 2.9</td><td> 2.9</td><td> 2.9</td><td> 2.9</td><td> 2.9</td>

n.d. = not determined

Table 9 - Accelerated (55’C) stability data of 2 mg/ml epirubicin.HCl solutions in 5% Dextrose at various pHs.

Time (hours)

Buffers Tests 0 4 8 16 24 34 48 72 _96___120 __144__

Epirubicin.HCl assay pH 2.5 . ng/ml 2.105 1.921 1.909 1.815 1.819 1.624 1.521 1.264 glycine.HCl . I initial 100.0 n.d. n.d. 91.3 90.7 86.2 86.4 77.1 72.3 n.d. 60.0 pH 2.4 2.3 2.3 2.3 2.3 2.3 2.3 2.3

Epirubicin.HCl assay

<td> pH 3.0</td><td> . mg/ml</td><td> 2.029</td><td> 1.990 1.914 1.949</td><td> 1.866 1.743</td><td> 1.562</td><td> 1.442 1.318</td>

<td> glycine.HCl</td><td> . 1 initial</td><td> 100.0</td><td> n.d. 98.1 94.3 96.1</td><td> 92.0 85.9</td><td> n.d. 77.0</td><td> 71.1 65.0</td>

<td></td><td> pH</td><td> 2.9</td><td> 2.8 2.9 2.8</td><td> 2.8 2.8</td><td> 2.8</td><td> 2.8 2.8</td>

n.d. = not determined

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Table 10 - K_obs values (1/days) for the degradation of epirubicin.HC1 solutions in water for Injection at various pHs at 55’C.

Buffer pH K״_ks χ 10³ 95% confidence limits ^c ODS

<td> Glycine.HC1</td><td></td><td></td><td></td>

<td> (0.05 ־־ I)</td><td> 2.5</td><td> 55.1</td><td> + 4.0</td>

Glycipe-HCl (I = 0.05)

3.0

66.8 ± 5.4

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Table 10 - K_obs values (1/days) for the degradation of epirubicin.HCl solutions in water for Injection at various pHs at 55°C.

<td> Buffer</td><td> PH</td><td> Κ־_Μ x I»³</td><td> 95% confidence limits</td>

<td> Glycine.HC1 (I = 0.05)</td><td> 2.5</td><td> 55.1</td><td> ± 4.0</td>

<td> Glycine-HCl (I = 0.05)</td><td> 3.0</td><td> 66.8</td><td> ± 5.4</td>

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Table 11 K_obs values (1/days) for the degradation of epirubicin.HCl 2 mg/ml solutions in 0.9% Sodium Chloride Injection at various pHs at 55‘C.

Buffer

PH ^Kobs x 10³

95% confidence limits

Glycine.HC1 (I = 0.05)

2.5 97.3 ± 6.4

Glycine-HCl (I = 0.05)

3.0

84.8 ± 6.5

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Table 12 - K_obs values (1/days) for the degradation of epirubicin.HCl 2 mg/ml solutions in 5% Dextrose solution at various pHs at 55°C.

<td> Buffer</td><td> PH</td><td> K־b_S^x 1°’</td><td> 95% confidence limits</td>

<td> Glycine.HC1 (I = 0.05)</td><td> 2.5</td><td> 81.1</td><td> ± 6.6</td>

<td> Glycine-HCl (I = 0.05)</td><td> 3.0</td><td> 70.9</td><td> ± 4.8</td>

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