(2S,3R,4S,5S,6R)-4-amino-2-[(1S,2S,3R,4S,6R)-4,6-diamino-3-[(2R,3R,5S,6R)-3-amino-6-(aminomethyl)-5-hydroxyoxan-2-yl]oxy-2-hydroxycyclohexyl]oxy-6-(hydroxymethyl)oxane-3,5-diol

Conditions: temp:0.00; pressure:0.00; stir:; See reaction.notes.procedure_details.

Procedure: Foam wound materials containing tobramycin sulfate were prepared as shown in Example 1. Once the materials were placed onto grid supports they were maintained at room temperature and covered with a foil wrap. Every 12 hours for nine days, 5 ml of water was applied onto the foam bandage materials and eluent was collected. Two ml samples containing SDMC-entrapped tobramycin were assayed for tobramycin antibiotic activity using standard antimicrobial assays. The results are shown in Table IX. were prepared was collected Two ml samples containing SDMC-entrapped tobramycin

Conditions: temp:0.00; pressure:0.00; stir:; See reaction.notes.procedure_details.

Procedure: To a solution of 50 ml of dichloromethane was added 10 grams of soy lecithin and 200 mg of tobramycin sulfate in 5 ml of water. Fifty ml of additional dichloromethane was added and the solution was mixed by shaking. Eight ml of solution (4 ml per side) was added to a 1"×1" foam wound bandage material which was then allowed to air dry. The materials were analyzed for residual dichloromethane by high pressure liquid chromatography and found to contain no detectable residual dichloromethane. The 1"×1" foam wound materials were packaged individually in heat sealed bags and stored at room temperature for two months. At the end of two months the packages were opened and a representative first sample of SDMC-forming foam wound bandages were removed, placed onto grid supports, and wetted with 5 ml of water. Two ml of eluent containing SDMC-entrapped tobramycin were collected from the first sample. A second representative sample of SDMC-forming foam wound bandages were stressed by heating to +70° C. for three days. A third representative sample of SDMC-forming foam wound bandages were stressed by freezing at -70° C. for three days, then allowed to equilibrate to room temperature. These stressed foam wound dressings were then placed onto grid supports, wetted with 5 ml of water, and 2 ml of eluent containing SDMC-entrapped tobramycin was collected. All samples were subjected to analysis for tobramycin entrapped as explained in Example 3. The results are shown in Table VII. the solution was mixed to air dry in heat sealed bags stored at room temperature for two months At the end of two months were removed Two ml of eluent containing SDMC-entrapped tobramycin were collected from the first sample by heating to +70° C. for three days to equilibrate to room temperature containing SDMC-entrapped tobramycin was collected

Conditions: temp:0.00; pressure:0.00; stir:; See reaction.notes.procedure_details.

Procedure: To a solution of 75 ml of dichloromethane was added 10 grams of soy lecithin (with tracer amounts of 3 -Hphosphatidlylcholine) and 200 mg of tobramycin sulfate in 5 ml of water. The solution was mixed by shaking and 4 ml were applied onto each side of 1"×1" foam bandage material and allowed to air dry. The bandage materials were placed onto a grid support as in Example 1 and wetted with 5 ml of water. The eluents for the first nine wettings were discarded. After ten repeated wettings the eluent was collected and passed over a Sephadex G-150 column. The material which was excluded by the column and included by the column bed was assayed for tobramycin using standard antimicrobial assays with E. Coli strain Y-1089. The material which was excluded was subjected to turbidity measurements and laser light scattering analysis. The sizes of the SDMCs were consistent with those previously observed between 0.15 m and 0.3 m and when assayed for tobramycin demonstrated that the antibiotic was indeed entrapped within the SDMCs which eluted from the foam bandage material. The included volume contained a small amount (less than 10% of the total tobramycin) which had either not been entrapped in SDMCs or had leaked out of SDMCs during transit from foam dressing or over the Sephadex column. The solution was mixed to air dry After ten repeated wettings the eluent was collected eluted from the foam bandage material

Conditions: temp:0.00; pressure:0.00; stir:; See reaction.notes.procedure_details.

Procedure: To a solution of 75 ml of dichloromethane was added 10 grams of soy lecithin and 200 mg of tobramycin sulfate in 5 ml of water. The solution was mixed by shaking and 4 ml were applied onto each side of the 1"×1" foam bandage material and allowed to air dry. Four ml of a solution comprising 20 mg of tobramycin sulfate in 8 ml of water was applied onto each side of 10 additional foam bandage material pieces (1"×1") and allowed to air dry. The foam bandage pieces were placed onto a grid support, wetted with 5 ml of water and 2 ml of eluent was collected which flowed through the foam dressing and into the underlying receptacle. The foam dressings were wetted subsequent times and 2 ml of eluent was collected. The antimicrobial activity of eluent was determined using standard antimicrobial inhibition assays using E. coli strain Y-1089. The results as shown in Table II. The solution was mixed to air dry to air dry was collected which was collected The results

Conditions: temp:0.00; pressure:0.00; stir:; See reaction.notes.procedure_details.

Procedure: When the requisite 5-O-thioformyl intermediate is prepared by reacting tobramycin with an excess of benzoyl chloroformate and potassium carbonate in aqueous methanol in a manner similar to that described in Preparation 1A(1) followed by reaction of the thereby formed 1,3,2',6',3"-penta-N-benzyloxycarbonyltobramycin with benzoyl chloride in pryidine in the manner of Preparation 1C(1) thence reaction of the resulting 1,3,2',6',3"-penta-N-benzyloxycarbonyl-4',2",4",6"-tetra-O-benzoyltobramycin with N,N-dimethyl-α-chloroformimidinium chloride in methylene chloride in pyridine followed by treatment with dry hydrogen sulfide in a manner similar to that described in Preparation 1D(1) whereby is obtained 1,3,2',6',3"-penta-N-benzloxycarbonyl-5-O-thioformyl-4',2",4",6"-tetra-O-benzoyltobramycin. When the requisite 5-O-thioformyl intermediate is prepared that described in Preparation followed by reaction of the

Conditions: temp:0.00; pressure:0.00; stir:; See reaction.notes.procedure_details.

Procedure: Fosfomycin disodium (18.057 g, 13.99 g free acid) was dissolved in 250 mL of water and the pH was adjusted to 7.41 by the dropwise addition of 1.53 mL of 4.5 N HCl. To the resulting solution was added 1.56 g of 97.5% tobramycin base. The pH of the solution was adjusted to 7.60 by the addition of 2.45 mL of 4.5 N HCl. The solution was diluted to 500 mL with water and filtered through a 0.2 μm Nalge Nunc 167-0020 membrane filter. The final pH was 7.76, the osmolality was 537 mOsmol/kg, the fosfomycin/tobramycin ratio was calculated to be 9:1, and the chloride concentration was 35.8 mM. filtered through a 0.2 μm Nalge Nunc 167-0020 membrane filter

Conditions: temp:0.00; pressure:0.00; stir:; See reaction.notes.procedure_details.

Procedure: Fosfomycin disodium (18.057 g, 13.99 g free acid) was dissolved in 250 mL of water and the pH was adjusted to 7.41 by the dropwise addition of 1.53 mL of 4.5 N HCl. To the resulting solution was added 1.56 g of 97.5% tobramycin base. The pH of the solution was adjusted to 7.60 by the addition of 2.45 mL of 4.5 N HCl. The solution was diluted to 500 mL with water and filtered through a 0.2 μm Nalge Nunc 167-0020 membrane filter. The final pH was 7.76, the osmolality was 537 mOsmol/kg, the fosfomycin/tobramycin ratio was calculated to be 9:1, and the chloride concentration was 35.8 mM. filtered through a 0.2 μm Nalge Nunc 167-0020 membrane filter

Conditions: temp:0.00; pressure:0.00; stir:; See reaction.notes.procedure_details.

Procedure: Foam wound materials containing tobramycin sulfate were prepared as shown in Example 1. Once the materials were placed onto grid supports they were maintained at room temperature and covered with a foil wrap. Every 12 hours for nine days, 5 ml of water was applied onto the foam bandage materials and eluent was collected. Two ml samples containing SDMC-entrapped tobramycin were assayed for tobramycin antibiotic activity using standard antimicrobial assays. The results are shown in Table IX. were prepared was collected Two ml samples containing SDMC-entrapped tobramycin

Conditions: temp:0.00; pressure:0.00; stir:; See reaction.notes.procedure_details.

Procedure: To a solution of 50 ml of dichloromethane was added 10 grams of soy lecithin and 200 mg of tobramycin sulfate in 5 ml of water. Fifty ml of additional dichloromethane was added and the solution was mixed by shaking. Eight ml of solution (4 ml per side) was added to a 1"×1" foam wound bandage material which was then allowed to air dry. The materials were analyzed for residual dichloromethane by high pressure liquid chromatography and found to contain no detectable residual dichloromethane. The 1"×1" foam wound materials were packaged individually in heat sealed bags and stored at room temperature for two months. At the end of two months the packages were opened and a representative first sample of SDMC-forming foam wound bandages were removed, placed onto grid supports, and wetted with 5 ml of water. Two ml of eluent containing SDMC-entrapped tobramycin were collected from the first sample. A second representative sample of SDMC-forming foam wound bandages were stressed by heating to +70° C. for three days. A third representative sample of SDMC-forming foam wound bandages were stressed by freezing at -70° C. for three days, then allowed to equilibrate to room temperature. These stressed foam wound dressings were then placed onto grid supports, wetted with 5 ml of water, and 2 ml of eluent containing SDMC-entrapped tobramycin was collected. All samples were subjected to analysis for tobramycin entrapped as explained in Example 3. The results are shown in Table VII. the solution was mixed to air dry in heat sealed bags stored at room temperature for two months At the end of two months were removed Two ml of eluent containing SDMC-entrapped tobramycin were collected from the first sample by heating to +70° C. for three days to equilibrate to room temperature containing SDMC-entrapped tobramycin was collected

Conditions: temp:0.00; pressure:0.00; stir:; See reaction.notes.procedure_details.

Procedure: To a solution of 75 ml of dichloromethane was added 10 grams of soy lecithin (with tracer amounts of 3H-phosphatidlylcholine) and 200 mg of tobramycin sulfate in 5 ml of water. The solution was mixed by shaking and 4 ml were applied onto each side of 1"×1" foam bandage material and allowed to air dry. The bandage materials were placed onto a grid support as in Example 1 and wetted with 5 ml of water. The eluents for the first nine wettings were discarded. After ten repeated wettings the eluent was collected and passed over a Sephadex G-150 column. The material which was excluded by the column and included by the column bed was assayed for tobramycin using standard antimicrobial assays with E. Coli strain Y-1089. The material which was excluded was subjected to turbidity measurements and laser light scattering analysis. The sizes of the SDMCs were consistent with those previously observed between 0.15 μm and 0.3 μm and when assayed for tobramycin demonstrated that the antibiotic was indeed entrapped within the SDMCs which eluted from the foam bandage material. The included volume contained a small amount (less than 10% of the total tobramycin) which had either not been entrapped in SDMCs or had leaked out of SDMCs during transit from foam dressing or over the Sephadex column. The solution was mixed to air dry After ten repeated wettings the eluent was collected eluted from the foam bandage material

Conditions: temp:0.00; pressure:0.00; stir:; See reaction.notes.procedure_details.

Procedure: To a solution of 75 ml of dichloromethane was added 10 grams of soy lecithin and 200 mg of tobramycin sulfate in 5 ml of water. The solution was mixed by shaking and 4 ml were applied onto each side of the 1"×1" foam bandage material and allowed to air dry. Four ml of a solution comprising 20 mg of tobramycin sulfate in 8 ml of water was applied onto each side of 10 additional foam bandage material pieces (1"×1") and allowed to air dry. The foam bandage pieces were placed onto a grid support, wetted with 5 ml of water and 2 ml of eluent was collected which flowed through the foam dressing and into the underlying receptacle. The foam dressings were wetted subsequent times and 2 ml of eluent was collected. The antimicrobial activity of eluent was determined using standard antimicrobial inhibition assays using E. coli strain Y-1089. The results as shown in Table II. The solution was mixed to air dry to air dry was collected which was collected The results

Conditions: temp:0.00; pressure:0.00; stir:; See reaction.notes.procedure_details.

Procedure: Foam wound materials containing tobramycin sulfate were prepared as shown in Example 1. Once the materials were placed onto grid supports they were maintained at room temperature and covered with a foil wrap. Every 12 hours for nine days, 5 ml of water was applied onto the foam bandage materials and eluent was collected. Two ml samples containing SDMC-entrapped tobramycin were assayed for tobramycin antibiotic activity using standard antimicrobial assays. The results are shown in Table IX. were prepared was collected Two ml samples containing SDMC-entrapped tobramycin