MEDICAL COMPOSITIONS AND THEIR USE IN TREATING PANCREATIC CANCER

Abstract

The invention relates to a pharmaceutical composition comprising (i) a live strain from the genus of Kazachstania spp, Acetobacter spp. and/or Stenotrophomonas spp, (ii) an attenuated strain from the genus of Kazachstania spp, Acetobacter spp. and/or Stenotrophomonas spp or, (iii) a medically active substance isolated from a strain from the genus of Kazachstania spp, Acetobacter spp. and/or Stenotrophomonas spp. These may be used to treat pancreatic cancer.

Claims

1. A pharmaceutical composition comprising: (iv) a live strain from the genus of Kazachstania spp, Acetobacter spp, and/or Stenotrophomonas spp for use in treating a pancreatic cancer, or a cancer associated with an infection or dysbiosis with Malassezia spp, (v) an attenuated strain from the genus of Kazachstania spp, Acetobacter spp. and/or Stenotrophomonas spp for use in treating pancreatic cancer or a cancer associated with an infection or dysbiosis with Malassezia spp or, (vi) a medically active substance isolated from a strain from the genus of Kazachstania spp, Acetobacter spp. and/or Stenotrophomonas spp for use in treating pancreatic cancer or a cancer associated with an infection or dysbiosis with Malassezia spp.

2. A pharmaceutical composition according to claim 1, wherein the strain is selected from the group of Kazachstania unispora, Acetobacter fabarum and Stenotrophomonas maltophila.

3. A pharmaceutical composition according to claim 1, wherein the composition inhibits growth and/or cell division of Malassezia spp.

4. A pharmaceutical composition according to claim 1, wherein the pancreatic cancer is a pancreatic ductal andenocarcinoma and/or wherein it is associated with an infection or dysbiosis with Malassezia spp.

5. A pharmaceutical composition according to claim 1, wherein the cancer is selected from the group of pancreatic ductal adenocarcinoma, small intestine cancer, large intestine cancer, gallbladder cancer, and liver cancer.

6. A microbial strain selected from the genus of Kazachstania spp, Acetobacter spp. and/or Stenotrophomonas spp for use as a medicament, (i) a cellular extract from a strain selected from the genus of Kazachstania spp, Acetobacter spp. and/or Stenotrophomonas spp for use as a medicament in treating pancreatic cancer associated with an infection or dysbiosis with Malassezia spp, (ii) the supernatant from the cell culture from a strain selected from the genus of Kazachstania spp, Acetobacter spp. and/or Stenotrophomonas spp for use as a medicament in treating pancreatic cancer associated with an infection or dysbiosis with Malassezia spp or, (iii) a medically active molecule or a medically active substance obtained from a strain selected from the genus of Kazachstania spp, Acetobacter spp. and/or Stenotrophomonas spp for use as a medicament in treating pancreatic cancer associated with an infection or dysbiosis with Malassezia spp.

7. A strain according to claim 6, wherein the strain is selected from the group of Kazachstania unispora, Acetobacter fabarum and Stenotrophomonas maltophila.

8. A strain, extract, supernatant, molecule or substance according to claim 6, wherein the strain, extract, supernatant, molecule or substance inhibits growth and/or cell division of Malassezia spp.

9. A strain, extract, supernatant, molecule or substance according to claim 6, wherein the pancreatic cancer is a pancreatic ductal andenocarcinoma.

10. A strain, extract, supernatant, molecule or substance according to claim 6, wherein the cancer is selected from the group of pancreatic ductal adenocarcinoma, small intestine cancer, large intestine cancer, gallbladder cancer, and liver cancer.

Description

FIGURE CAPTIONS

[0104] FIG. 1|Inhibition assays of the fungicide ketoconazole as positive control for inhibition and Kazachstania unispora supernatant on M. sympodialis, M. furfur, M. restricta and M. globosa. Inhibition assays were performed using the disc diffusion method. Discs were soaked with ketoconazole (1 mg/ml) or K. unispora supernatant and placed on agar plates containing a soft layer inoculated with a single culture of the different Malassezia species to be tested. On the left column of pictures, halo with no growth indicates that ketoconazole inhibits growth of all 4 Malassezia species tested. On the right column, an inhibition halo is only seen on M. restricta and M. globosa agar plates, indicating that K. unispora inhibits growth of those two species but that it does not inhibit growth of M. sympodialis and M. furfur.

[0105] FIG. 1|Inhibition assays of M. restricta and M. globose by growth of Acetobacter fabarum. Discs were soaked with ketoconazole (1 mg/ml in DMSO) or exponentially growing Acetobacter fabarum culture and placed on agar plates containing a soft layer inoculated with a single culture of the different Malassezia species to be tested. In the upper panel, inhibition of Malassezia globosa is shown; in the bottom panel, Malassezia restricta is shown. The halo with no growth indicates that ketoconazole inhibits growth of both Malassezia species tested. An inhibition halo is also seen around the disk imbued with growing culture of Acetobacter fabarum for both species of Malassezia tested.

EXAMPLES

Example 1

[0106] In order to identify strains inhibiting Malassezia growth, inhibition assays were performed using the disc diffusion method. This method consists in disposing discs imbibed with potential inhibitory substances on Petri plates containing a strain of interest. After optimal growth of the strain of interest, growth inhibition can be observed around discs soaked with substances having an inhibitory effect. Potential inhibitory substances tested in this experiment are supernatants obtained after individually culturing a collection of fungal and bacterial strains. After an incubation of 10 days in their optimal broth, each culture was centrifuged and the supernatant was filtered and further stored at −20° C. Ketoconazole, a fungicide, was used as a positive control for Malassezia growth inhibition. It was diluted in dimethyl sulfoxide at 1 mg/ml. The collection of supernantants was tested on the following species from the genus Malassezia: Malassezia restricta, Malassezia globosa, Malassezia furfur and Malassezia sympodialis. All strains were streaked from −80° C. stock on Modified Leeming and Notman (MLN) agar plates and incubated 10 days at 30° C. A 5 ml liquid culture of each species was made in MLN broth and incubated at 30° C. for 2 days for M. furfur and M. sympodialis and 5 days for M. restricta and M. globosa. To allow a lawn growth of the different Malassezia strains tested, Petri plates were prepared as follows: a bottom layer of 50 ml Modified Sabouraud Dextrose (MSD) agar was poured in Petri plates. The soft agar, constituting the top layer, was a mix of 12.5 ml MSD agar, 10.5 ml MLN broth and 2 ml of liquid culture. Plates were allowed to dry for one hour, then, 6 mm-diameter Whatman™ discs were soaked with 15 ul of each thawed supernatants and ketoconazole and placed on the soft agar plate containing the solidified Malassezia monoculture layer. After an incubation of 5 days, inhibition area could be easily observed around discs. As expected, ketoconazole inhibited all 4 Malassezia species tested in this experiment. In addition, one supernantant was able to inhibit the growth of Malassezia globosa and Malassezia restricta while having no effect on Malassezia furfur and Malassezia sympodialis (see FIG. 1). The strain from which the inhibitory supernatant has been produced was further sequenced using the Sanger technology. The internal transcribed spacer (ITS) of this strain was taxonomically assigned to the yeast Kazachstania unispora (BLASTn, 99% identity, 99% query cover, e-value 0.00).

Example 2

[0107] In order to identify strains whose growth inhibits Malassezia, inhibition assays were performed using a modified disc diffusion method. This method is similar to the classical disk diffusion method that consists in depositing discs imbibed with potential inhibitory substances on Petri plates containing a strain of interest, except that the disks were imbibed with a living culture of an Acetobacter strain. After optimal growth of the strain of interest, growth inhibition can be observed around discs on which the growth of inhibitory bacteria has occurred. Ketoconazole (at 1 mg/mL in DMSO) was used as a positive control for Malassezia growth inhibition. The effect of growth of Acetobacter was tested on Malassezia restricta and Malassezia globosa. All Malassezia strains were streaked from −80° C. stock on Modified Leeming and Notman (MLN) agar plates and incubated 10 days at 30° C. A 5 ml liquid culture of each species was made in MLN broth and incubated at 30° C. for 5. To allow a lawn growth of the different Malassezia strains tested, Petri plates were prepared as follows: a bottom layer of 50 ml Modified Sabouraud Dextrose (MSD) agar was poured in Petri plates. The soft agar, constituting the top layer, was a mix of 12.5 ml MSD agar, 10.5 ml MLN broth and 2 ml of liquid culture. Plates were allowed to dry for one hour, then, 6 mm-diameter Whatman™ discs were soaked with 15 ul of exponentially growing cultures of Acetobacter and placed on the soft agar plate containing the solidified Malassezia monoculture layer. After an incubation of 5 days, inhibition area could be easily observed around discs (see below). As expected, ketoconazole inhibited both Malassezia restricta and Malassezia globose. In addition, one supernantant was able to inhibit the growth of Malassezia globosa and Malassezia restricta (see FIG. 2).