USE OF OLIGONUCLEOTIDES FOR THE TREATMENT OF TUMORS

Abstract

In a method of treating a patient having a solid tumor, the solid tumor is completely or partially resected or ablated and then a therapeutically-effective amount of an oligonucleotide that kills tumor cells is applied or administered in a body cavity created by the resection or ablation in order to kill tumor cells remaining in, or in the surroundings of, a tumor bed and/or to suppress metastases. Thus, the oligonucleotide counteracts the development of recurrences of the solid tumor or new metastases. The oligonucleotide may act, e.g., in a pleiotropic manner in at least one type of tumor cell.

Claims

1. A method of treating a patient having a solid tumor, comprising: performing a surgical procedure and creating an incision in the patient to perform complete or partial resection or ablation of the solid tumor, and then, before completing the surgical procedure and closing the incision or before removing a drainage inserted during the surgical procedure, locally applying a therapeutically-effective amount of an oligonucleotide that kills tumor cells in a body cavity created by the resection or ablation in order to fight tumor cells remaining in, or in the surroundings of, a tumor bed and/or to fight metastases, the oligonucleotide counteracting development of recurrences of the solid tumor or new metastases.

2. The method according to claim 1, wherein the oligonucleotide acts in a pleiotropic manner in at least one type of tumor cell.

3. The method according to claim 1, wherein the oligonucleotide is applied in a gel, in a liquid or in an elastic material.

4. The method according to claim 1, wherein the oligonucleotide is applied as a load of a carrier in the form of a gel, a liquid or an elastic material, and wherein the carrier contains the oligonucleotide at a concentration of more than 3 micrograms per milliliter.

5. The method according to claim 4, wherein the carrier comprises a resorbable elastic or gel material and at least partially reproduces the shape of the resected solid tumor.

6. The method according to claim 1, wherein the oligonucleotide is applied as a load of a flat carrier composed of a membrane, a foil or a gauze, wherein the oligonucleotide is carried in the flat carrier directly, packaged in particles, conjugated thereto or formulated as a gel, and wherein that the oligonucleotide is present in the flat carrier in a loading density of at least 10 micrograms per square centimeter.

7. The method according to claim 4, wherein the carrier contains collagen, atelocollagen, gelatin, chitosan or hyaluronic acid.

8. The method according to claim 6, wherein the carrier additionally contains one or more absorptive or fluorescent dyes.

9. The method according to claim 4, wherein the carrier comprises a crosslinked or otherwise stabilized material that degrades in the patient's body at an in vivo half-life of more than a week.

10. The method according to claim 4, wherein the carrier further comprises a therapeutically-effective amount of a chemotherapy medicament.

11. The method according to claim 4, wherein the carrier is composed of two or more of the gel, the gauze and the liquid to optimally reach tumor cells which are located at a distance from a wound margin created by the resection or ablation.

12. The method according to claim 4, further comprising one or more diagnostic elements integrated on the carrier.

13. The method according to claim 1, further comprising carrying out at least one standard therapy selected from aftercare and adjuvant chemotherapy in parallel with or almost simultaneously with the the steps of tumor resection or ablation and oligonucleotide application.

14. The method according to claim 1, wherein the method is for adjuvant therapy of adrenocortical carcinoma, ovarian cancer, mesothelioma or glioblastoma.

15. The method according to claim 1, wherein the oligonucleotide is applied locally to the tumor bed or to a surgical margin in a carrier that has an initial elution rate of more than 1 microgram/square centimeter per day.

16. The method according to claim 1, wherein the oligonucleotide is applied laparoscopically to the tumor bed or to a surgical margin using a brush.

17. The method according to claim 1, wherein the oligonucleotide is applied as an aerosol to the tumor bed or to a surgical margin using a brush.

Description

PREFERRED EMBODIMENTS

[0048] A treatment using only one application is preferred, since access to the surgical cavity is limited in time, in view of the rapid development of laparoscopic surgical techniques used in the case of tumor resections, and, ideally, currently does not exceed more than 30 minutes. Each subsequent opening of the surgical cavity would also increase the risk of disseminating tumor cells.

[0049] According to the invention, preferably pleiotropic oligonucleotides are used in order to act on as many tumor cells as possible, irrespective of the phase of the cell cycle in which the cell is currently located, in particular in the case of a single application. According to the invention, pleiotropic oligonucleotides are those oligonucleotides which act simultaneously in a cell against at least two targets, for example against two mRNAs having the same target sequence, which mRNAs code for different proteins. Prodrugs according to WO 2010/102615 are an example for a pleiotropic oligonucleotide, wherein preferred embodiments may be effective in tumor cells against several physiological targets in parallel (WO 2012/098234).

[0050] According to the invention, preferred is the use of biodegradable carrier materials having low immunogenicity, which exhibit hydrogel-like or elastic properties, and which elute oligonucleotides by diffusion or during in vivo degradation or by changes in parameters of the surroundings such as pH. Carriers, which are medically proven materials such as collagen, atelocollagen, gelatin, fibrin, chitosan or hyaluronic acid, synthetic or recombinant variants thereof and their synthetic modifications, are used with particular preference.

[0051] It is particularly preferred if the use of oligonucleotides according to the invention is in carriers which are temporarily associated, such that a large amount of oligonucleotide is available locally and over an extended period of time. According to the invention, “temporarily associated” means that the carrier may be solid, elastic or even deformable, but constitutes a unit over several hours or days. Biodegradable carriers also are considered to be temporarily associated, wherein the carriers become smaller as a result of breakdown processes within the body and which disintegrate into smaller units after hours or days, which are dismantled from a carrier like a gauze, or which undergo a transformation into a liquid phase.

[0052] The use of the oligonucleotides according to the invention is particularly preferred in large amounts per application. The total amount of oligonucleotides used depends on the resulting surgical cavity and the structure of the surrounding tissue. For the associated carriers, this association is described as loading density. The use of oligonucleotides is preferred in carriers with loading densities of more than 3 micrograms of oligonucleotide per milliliter of carrier volume, particularly preferably more than 12, 50, 250, 1000 or 5000 micrograms per milliliter, respectively.

[0053] The use of carriers may also be advantageous, from which the oligonucleotides elute over an extended period of time, in particular over several days. Carriers that are absorbed by the body are particularly advantageous. Carriers that make possible a simple laparoscopic handling, such as flexible foils or rods, are particularly advantageous. Oligonucleotides and smaller particulate carriers may be applied in nets, such as nets made of gauze material, or in a hydrogel. Preferred is the use of carriers which have an initial elution rate of more than 1 microgram/square centimeter and day, in particular more than 2, 5, 10, 25 or 100 microgram/square centimeter and day, respectively.

[0054] According to the invention, the oligonucleotides are used locally after the tumor resection in order to inhibit a recurrence-development or metastases-development. Accordingly, after the complete or partial removal of the primary tumor, they are applied during the surgery to the resulting surgical margins and the surrounding tissue. This also includes surrounding connective and fatty tissues. For a mechanical application of the oligonucleotides, gel-like formulations are advantageous, which may be distributed or sprayed onto the tissue with a brush or similar tool.

[0055] Collagen, which may be provided with gel-like or elastic properties, is suitable as a carrier material in the use of the oligonucleotides. Collagen is absorbed by the body and exhibits low immunogenicity.

[0056] The use of oligonucleotides in liquid formulation according to the invention is advantageous if the oligonucleotides may be distributed in the surgical cavity using a syringe. An advantage of this form of application is provided in the case of complex shapes of surgical cavities because the formulation distributes itself within the surgical cavity and may flow out of its openings, just as likely as seeding metastases. “Complex” in the sense of the invention means that the surgical cavity is open to cavities such as the peritoneum, through which tumor cells can get into remote margins of the peritoneum.

[0057] A particularly suitable distribution is ensured by the use of liquid volumes that are of similar size, particularly preferably of the same size or even 1, 2, 5 times as large as the volume of the surgical cavity alone or including cavities connected thereto. For example, in the case of the removal of an adrenal gland, a surgical cavity of approximately 25 ml is created, which is connected, however, to the much larger retroperitoneal space. With large volumes of liquid, it is possible to also carry out perfusions, such that tumor-effective oligonucleotides are continuously supplied to the surgical cavity and high local concentrations of oligonucleotides may be achieved everywhere in the cavity.

[0058] The use of oligonucleotides according to the invention as an aerosol is also preferred. For this purpose, an overpressure is created in the surgical cavity or in the cavities adjacent thereto. The aerosol is introduced into the resulting cavity through a trocar, tube or similar tool or is even generated with an inserted aerosol generator. The aerosol is distributed almost uniformly in the cavity, such that even remote areas of the cavity are reached. The cavity is maintained open for more than two hours, and particularly more than an hour or half an hour. The administration of the oligonucleotides according to the invention in an aerosol form may also be repeated several times if access to the cavity is continuous or may be re-established quickly and easily.

[0059] Preferred is the use of oligonucleotides in aerosols in amounts of more than 1 microgram/square centimeter of cavity volume, and particularly preferably of more than 2.5, 10, 25, 100 micrograms.

[0060] The use of the oligonucleotides according to the invention includes combinations with methods and therapies known from the prior art. This includes, for example, the simultaneous use of medicaments such as cytostatics and oligonucleotides, and also, for example, a radiotherapy carried out in parallel.

EXEMPLARY EMBODIMENT

Adrenocortical Carcinoma

[0061] Carcinomas of the adrenal cortex are rare in occurrence and as of today are usually treated by laparoscopic adrenalectomy, i.e., the minimally invasive surgical removal of the complete adrenal gland. Prognosis for patients thus treated nevertheless is poor, particularly in the case of late-identified primary tumors; recurrences or metastases are very likely to occur and usually lead to the patient's death within a few months.

[0062] Laparoscopic adrenalectomy is performed using either abdominal/transperitoneal or retroperitoneal access. The renal fascia is pierced and a surgical cavity is created, which is pressurized at 20-30 mm Hg positive pressure for stabilization. The positive pressure also reduces or prevents bleeding into the surgical cavity following any injury to blood vessels. Subsequently, further incisions are used to mobilize surrounding organs, to sever arteries and veins supplying the adrenal gland, and to expose the adrenal gland. Ideally, the adrenal gland as a whole is dissected together with the surrounding adipose tissue, transferred to a retrieval pouch, which in turn is pulled out of the patient via one of the trocars. A lymphadenectomy may be performed in addition. The surgical cavity is rinsed with distilled water and antibiotics and, in simple cases, closed without drainage.

[0063] In this surgical procedure, it is carefully avoided that the tumor capsule is damaged and that any tumor tissue remains in the patient. Nevertheless, local recurrences of unclear origin commonly occur. Tumor cells may have been transported from the adrenal gland into the surroundings prior to surgery. This includes seeding metastases occurring in body cavities on other organs or parts of organs caused by caudal migration of detached tumor cells by gravity. These processes are facilitated by the fact that the renal fascia surrounding the kidney opens medially and caudally into the retroperitoneal space.

[0064] According to the invention, oligonucleotides are used locally after tumor resection in order to prevent recurrence-development or metastases-development and to fight remaining tumor cells or metastases. To this end, for example, after removal of the adrenal gland, the oligonucleotides are applied during surgery to the resulting surgical margins and the surrounding tissue. This also includes surrounding connective and fatty tissue, especially the renal fascia. Particularly suitable for this purpose are gel-like formulations which may be applied laparoscopically to the wound margins using an instrument such as a brush. In the surgical cavity of an adrenocortical carcinoma, wound margins having a surface area of about 50 square centimeters are created. Gels are applied with a layer thickness of 0.2 to 1 mm, which corresponds to a gel volume of 1 to 5 ml. This gel volume preferably contains 70 micrograms of oligonucleotide and particularly preferably more than 250 or 1000 micrograms, or 5, 25 or 100 milligrams. The gel is degraded within a few weeks and the oligonucleotides are continuously released.

[0065] In a further exemplary embodiment, elastic carriers made of collagen, for example, are used, which are introduced into the surgical cavity. As highly elastic carriers, they are introduced into the surgical cavity via the aperture of a trocar as a capsule. After the encapsulation has been removed, the carrier takes on the size and shape corresponding to the resected organ of about 4*3*2 cm, having a volume or partial volume of less than 25 ml. It carries oligonucleotides in amounts of preferably more than 70 micrograms, and particularly preferably more than 250 or 1000 micrograms, or 5, 25 or 100 milligrams. The carrier is degraded within a few weeks and the oligonucleotides are continuously released.

[0066] The degradation rate of the gel and the elastic carrier is controllable by manufacturing parameters, in the case of collagen, for example, by the degree of cross-linking of the collagen. In vivo half-lives of more than 1 week are preferred, half-lives of 2 weeks, 4 weeks and 3 months are particularly preferred.

[0067] In a further embodiment according to the invention, a liquid formulation for the oligonucleotides is used. Here, the oligonucleotides are distributed in the surgical cavity, for example, by a syringe. An advantage of this form of application is that the formulation distributes itself within the surgical cavity and may flow out of its openings, just as likely as seeding metastases also would do. In adrenocortical carcinoma with the aforementioned expansion of the renal fascia, liquid forms of application reach its extensions much better than solid forms of application. The injected volume in this example is 20 ml, which comprises preferably more than 70 micrograms, and particularly preferred more than 250 or 1000 micrograms, or 5, 25 or 100 milligrams of oligonucleotide. The oligonucleotides may be packed in particles or liposomes such that they are released slowly only when the carriers are degraded or absorbed by the cells. The rate of degradation of the particles may be influenced by manufacturing parameters such as the degree of crosslinking. In vivo half-lives of more than 1 week are preferred, half-lives of 2 weeks, 4 weeks and 3 months are particularly preferred.

[0068] It may also be advantageous to use carriers having a simpler geometry, from which the oligonucleotides elute over an extended period, in particular over several days. Carriers which are absorbed by the body are particularly advantageous. Carriers which make possible a simple laparoscopic handling, such as flexible foils or rods, are particularly advantageous. Smaller particulate carriers may be applied in nets, such as gauze nets, or in a hydrogel.

[0069] In the case of adrenocortical carcinoma, the use of the oligonucleotides as an aerosol may be preferred, in particular in amounts of 70 micrograms, and particularly preferred in the amount of more than 250 or 1000 micrograms, or 5, 25 or 100 milligrams of oligonucleotide.

[0070] The combination of the adjuvant use of the oligonucleotides with adjuvant therapies such as mitotane and the combination with radiotherapy may be preferred.

EXEMPLARY EMBODIMENT

Ovarian Cancer

[0071] Currently, ovarian cancer is the sixth most common malignant disease in women (Guideline Ovarian Cancer 2013). The surgical removal of an ovary constitutes an essential part of the treatment. In the case of ovarian cancer, the result of the diagnosis strongly influences the extent in which tissue will be removed. In addition to the ovary itself, the fallopian tube and lymph nodes, and, in later stages, also other organs may be affected, in particular those extending into or adjacent to the peritoneum. The tumor resection is carried out as far as possible, but complete removal often is not possible in these cases because the surrounding tissue is already affected. In contrast to adrenocortical carcinoma, it is not recommended to perform the surgery laparoscopically.

[0072] Adjuvant chemotherapy is recommended in most cases; an exception to this recommendation relates to cases of very early stages at the time of surgery. Systemic application of carboplatin, a low molecular weight cytostatic, is recommended, which often is associated with significant side effects such as changes in blood count, dysfunction of the liver and nerves, as well as impaired cardiovascular function. In cases of a higher staging level, systemic therapy using paclitaxel and bevacizumab, a monoclonal antibody against VEGF, is also recommended. Relapses nevertheless occur frequently.

[0073] According to the invention, the oligonucleotides are used locally after the tumor resection in order to prevent recurrence-development or metastases-development and to fight remaining tumor cells or metastases. To this end, they are applied, for example, during surgery after removal of the ovary and other tissue to the resulting surgical margins and the surrounding tissue. This also includes surrounding connective and fatty tissue, especially parts of the peritoneum.

[0074] The size of an ovary in an adult is about 3.5*2*1 cm, its volume amounts to about 3-6 ml. Similar to adrenocortical carcinoma, various formulations are suitable, including a gel-like formulation. Despite the smaller organ volume compared to the adrenal cortex, volumes of 1-5 ml are used preferably since tissue surrounding the ovary should be coated to a greater extent. The gel volume preferably contains 70 micrograms and, particularly preferred, more than 250 or 1000 micrograms, or 5, 25 or 100 milligrams. The gel is degraded within a few weeks and the oligonucleotides are continuously released. Here, the oligonucleotides may be packed in particles or liposomes such that they are released slowly only when the carriers are degraded or are absorbed by the cells. The rate of degradation of the particles may be influenced by manufacturing parameters such as the degree of crosslinking. In vivo half-lives of more than 1 week are preferred, half-lives of 2 weeks, 4 weeks and 3 months are particularly preferred.

[0075] In a further exemplary embodiment, elastic carriers made of collagen, for example, are used, which are introduced into the surgical cavity and which completely or partially expand to the volume of an ovary. A carrier carries oligonucleotides in amounts of preferably more than 70 micrograms, and particularly preferred of more than 250 or 1000 micrograms, or 5, 25 or 100 milligrams. The carrier is degraded by the body within a few weeks and the oligonucleotides are continuously released.

[0076] The degradation rate of the gel and the elastic carrier are controllable by manufacturing parameters, in the case of collagen, for example, by the degree of cross-linking of the collagen. In vivo half-lives of more than 1 week are preferred, half-lives of 2 weeks, 4 weeks and 3 months are particularly preferred.

[0077] In cases of ovarian cancer, a liquid formulation for the oligonucleotides may also be advantageous, since the peritoneum surrounding the ovaries extends wide. The injected volume for ovarian cancer in this example is 20 ml, comprising preferably more than 70 micrograms, and particularly preferred more than 250 or 1000 micrograms, or 5, 25 or 100 milligrams of oligonucleotide. The oligonucleotides may be packed in particles or liposomes such that they are released slowly only when the carriers are degraded or are absorbed by the cells. The rate of degradation of the particles may be influenced by manufacturing parameters such as the degree of crosslinking. In vivo half-lives of more than 1 week are preferred, half-lives of 2 weeks, 4 weeks and 3 months are particularly preferred.

[0078] It may also be advantageous to use carriers having a simpler geometry, from which the oligonucleotides elute over an extended period, in particular over several days. Carriers which may be absorbed by the body, such as flexible foils or rods, are particularly advantageous. Smaller particulate carriers may be applied in nets, such as gauze nets, or in a hydrogel.

[0079] In the case of ovarian cancer, the use of the oligonucleotides as an aerosol may be preferred, in particular in amounts of 70 micrograms, and particularly preferred in the amount of more than 250 or 1000 micrograms, or 5, 25 or 100 milligrams of oligonucleotide.

[0080] The combination of the adjuvant use of the oligonucleotides with adjuvant cytostatics such as carboplatin, cisplatin, paclitaxel and bevacizumab, and the combination with radiation therapy may be preferred.

EXEMPLARY EMBODIMENT

Mesothelioma

[0081] Malignant diffuse mesothelioma is a tumor originating from the mesothelial or submesothelial cells of the pleura, peritoneum or pericardium. The prognosis for patients with malignant pleural mesothelioma is poor with median survival times of 4 to 12 months. A curative treatment is currently not available.

[0082] The predominate amount (>80%) of mesotheliomas originate from the pleura. Malignant mesotheliomas are comparatively rare. They mostly emerge as signal tumors of a previous exposure to asbestos (Neumann et al. 2013). It is also expected that cases of mesothelioma, e.g., after the attack on the World Trade Center in New York on September 11, 2001, will increase significantly in numbers during the next 15 years among those exposed at that time (Povtak 2016). Depending on the subtype of mesothelioma, in more than 50% of pleural mesotheliomas, tumor cells are released into the pleural effusion.

[0083] There is no standard therapy for treating mesothelioma. Recommended therapy concepts range from standalone symptomatic treatment to aggressive multimodal treatment comprising surgery, chemotherapy and radiation. Currently, two surgical strategies exist, namely, pleurectomy/decortication or extrapleural pleuropneumectomy, in order to achieve as complete a macroscopic tumor removal as possible. These resections are not performed laparoscopically. Due to the diffuse growth of the mesothelioma, however, a complete tumor removal generally is not possible. Residual parts of the tumor remain, which often are detectable only by microscope, in the case of which adjuvant radiotherapy and chemotherapy are recommended (Rice 2011). According to the prior art, a combination of intraperitoneal carboplatin and pemetrexed is recommended. Regarding patient survival, successes owing to the adjuvant use of check point inhibitors were recently noted (Scherpereel et al. 2017).

[0084] In contrast to carcinomas of the adrenal cortex and ovary, the size of the surgical cavity differs widely; after resection of a lung in pleural mesothelioma, it may amount to a size of several liters with an area of several hundred square centimeters. In the case of these volumes, a gel may also be used advantageously and applied with tools such as brushes; concentrations of more than 15 or 70 micrograms per milliliter of gel are preferred, particularly preferred of more than 250 or 1000 micrograms, or 5, 25 or 100 milligrams per milliliter of gel.

[0085] In the case of retaining the affected lung, a liquid formulation is suitable, preferably at concentrations of more than 15 or 70 micrograms per milliliter and particularly preferred at more than 250 or 1000 micrograms, or 5, 25 or 100 milligrams per milliliter.

[0086] In the case of large-volume resections, elastic carriers having a total volume corresponding to that of the resected organs are no longer useful. Instead, it is better that flat carriers, such as membranes, foils or gauze, are used, which carry oligonucleotides directly, packaged in particles such as liposomes, conjugated or formulated as a gel. Loading densities of 10 micrograms per square centimeter are preferred, particularly preferred are densities of 50 or 200 micrograms, or 1, 5 or 20 milligrams of oligonucleotide per square centimeter. Preferred is the application of oligonucleotides onto implants, such as those used after resection of the diaphragm, with the same oligonucleotide density.

[0087] Also in the case of mesothelioma, the use of carriers having a simpler geometry may be preferred, in particular if the oligonucleotides elute from these carriers over a longer period, in particular over several days. Carriers that are absorbed by the body, such as flexible foils or rods, are particularly advantageous. Smaller particulate carriers can be applied in nets, such as gauze nets, or in a hydrogel.

[0088] In mesothelioma, the use of the oligonucleotides as an aerosol may be preferred, in particular in amounts of 70 micrograms, and particularly preferred in the amount of more than 250 or 1000 micrograms, or 5, 25 or 100 milligrams of oligonucleotide.

[0089] The combination of the adjuvant use of the oligonucleotides with further adjuvant therapies such as carboplatin and pemetrexed, and the combination with radiotherapy may be preferred.

EXEMPLARY EMBODIMENT

Glioblastoma

[0090] Glioblastomas belong to the diffusely infiltrating, highly malignant gliomas and are the most common brain neoplasms with a share of 16%. According to the WHO classification, they are classified as grade IV tumors and are associated with a poor prognosis. Since glioblastomas exhibit a markedly infiltrative growth, a cure by surgical resection of the tumor is not possible. It is a goal to reduce the tumor mass surgically as completely as possible. An adjuvant therapy, in which radiotherapy is combined with chemotherapy, is therefore recommended according to the European Organization for Research and Treatment of Cancer and National Cancer Institute of Canada Clinical Trials Group (EORTC-NCIC) protocol (Javamanne et al., 2018). It commences the earliest at four weeks after the operation, however, when the surgical wound healing process has progressed. Currently, the treatment standard is temozolomide (Davis 2016), an oral alkylating chemotherapy drug, which is however genotoxic and teratogenic. The above-described adjuvant deposition of Gliadel™ wafers during the surgery is a treatment variant that is not part of the standard of treatment, but which also is carried out together with temozolomide treatment. In contrast, the simultaneous use of temozolomide and bevavizumab is not recommended (Holdhoff et al. 2011). A newer experimental treatment option includes DCVax, a procedure in which monocytes are removed from the patient and differentiated extracorporeally into dendritic cells carrying tumor antigens, and which are subsequently transferred back into the patient.

[0091] In glioblastoma, the size and shape of the surgical cavity differ individually. The size of the surgical cavity, however, also amounts to a few milliliters, making it comparable to that of adrenocortical carcinoma and ovarian carcinoma. The shape is different, however, and as individual as the size. In addition, partial resections are performed almost exclusively in the case of glioblastoma.

[0092] For the use according to the invention in the case of glioblastoma, gel-like formulations, which are applied to the wound margins using an instrument such as a brush, are also particularly suitable. The gel volume of 1 to 5 milliliters is preferred to contain 70 micrograms, particularly preferred more than 250 or 1000 micrograms, or 5, 25 or 100 milligrams. The gel is degraded within a few weeks and the oligonucleotides are continuously released. The oligonucleotides may be packed in particles or liposomes such that they are released slowly only when the carriers are degraded or are absorbed by the cells. The rate of degradation of the particles may be influenced by manufacturing parameters such as the degree of crosslinking. In vivo half-lives of more than 1 week are preferred, half-lives of 2 weeks, 4 weeks, 3 months or an entire year are particularly preferred.

[0093] In a further exemplary embodiment, elastic carriers made of collagen, for example, are used, which are introduced into the surgical cavity and which completely or partially expand to the volume of the surgical cavity. The one or the carriers carry oligonucleotides in amounts of preferably more than 70 micrograms, particularly preferred of more than 250 or 1000 micrograms, or 5, 25 or 100 milligrams. The one or the carriers are degraded by the body within a few weeks and the oligonucleotides are continuously released.

[0094] The degradation rate of a gel or of an elastic carrier is controllable by manufacturing parameters, in the case of collagen, for example, by the degree of cross-linking of the collagen. In vivo half-lives of more than 1 week are preferred, half-lives of 2 weeks, 4 weeks and 3 months are particularly preferred.

[0095] In cases of glioblastoma, a liquid formulation for the oligonucleotides may also be advantageous, since it is not known whether the glioblastoma also spreads in a manner other than infiltrating. In the case of glioblastoma, the injected volume, for example, amounts to 20 ml, in which preferably more than 70 micrograms, particularly preferred more than 250 or 1000 micrograms, or 5, 25 or 100 milligrams of oligonucleotide is found. The oligonucleotides may be packed in particles or liposomes such that they are released slowly only when the carriers are degraded or are absorbed by the cells. The rate of degradation of the particles may be influenced by manufacturing parameters such as the degree of crosslinking. In vivo half-lives of more than 1 week are preferred, half-lives of 2 weeks, 4 weeks and 3 months are particularly preferred.

[0096] In the case of glioblastoma, the use of the oligonucleotides as an aerosol may be preferred, in particular in amounts of 70 micrograms, and particularly preferred in the amount of more than 250 or 1000 micrograms, or 5, 25 or 100 milligrams of oligonucleotide

[0097] It may also be advantageous to use carriers having a simpler geometry, from which the oligonucleotides elute over an extended period, in particular over several days.

[0098] Carriers which are absorbed by the body, such as flexible foils or rods, are particularly advantageous. Smaller particulate carriers may be applied in nets, such as gauze nets, or in a hydrogel. Another example are wafers as used in Gliadel.

[0099] The combination of the adjuvant use of the oligonucleotides with adjuvant cytostatics such as temozolomide, with cellular therapies such as DCVax, and the combination with radiation may be preferred.

APPLICATION EXAMPLE

Peritoneal Carcinosis

[0100] Peritoneal carcinosis refers to the infestation of the peritoneum with multiple malignant tumor cells. The cause of a peritoneal carcinosis is usually not a tumor of the peritoneum itself, but rather a malignant tumor of another organ located in the abdomen. Most commonly, this is an advanced metastatic tumor of the gastrointestinal tract, of the pancreas or of the ovaries, as described above. In some cases, it is not possible to identify a primary tumor. (https://flexikon.doccheck.com/de/Peritonealkarzinose),

[0101] At 15-20 cases per 100,000 people, peritoneal carcinosis occurs rarely, but with an increasing tendency; in Germany there are about 35,000 new cases per year (Glockzin et al. 2007). The prognosis of the peritoneal carcinosis is on average about 6 months after diagnosis.

[0102] As long as it has not advanced too far, peritoneal cancer is being treated increasingly using multimodal therapy in which surgical cytoreduction is combined with an intraoperative, hyperthermal intraperitoneal chemotherapy (Piso et al. 2011). PIPAC, as described above, also is used.

[0103] In contrast to the adrenocortical and ovarian carcinomas, the size of the surgical cavity differs greatly individually due to the varying number of affected organs. A gel can nevertheless be advantageously used for these volumes and can be applied with tools such as brushes; concentrations of more than 15 or 70 micrograms per milliliter of gel are preferred, particularly preferred are concentrations of more than 250 or 1000 micrograms, or 5, 25 or 100 milligrams per milliliter of gel.

[0104] In the case of large-volume resections, elastic carriers having a total volume corresponding to that of the resected organs are no longer useful. Instead, it is better that flat carriers, such as membranes, foils or gauze, are used, which carry oligonucleotides directly, packaged in particles such as liposomes, conjugated or formulated as a gel. Loading densities of 10 micrograms per square centimeter are preferred, particularly preferred are densities of 50 or 200 micrograms, or 1, 5 or 20 milligrams of oligonucleotide per square centimeter. Preferred is the application of oligonucleotides onto implants, such as those used after resection of the diaphragm, with the same oligonucleotide density.

[0105] Also in the case of peritoneal carcinosis, the use of carriers having a simpler geometry may be preferred, in particular if the oligonucleotides elute from these carriers over a longer period, in particular over several days. Carriers that are absorbed by the body, such as flexible foils or rods, are particularly advantageous. Smaller particulate carriers can be applied in nets, such as gauze nets, or in a hydrogel.

[0106] In peritoneal carcinosis, the use of the oligonucleotides as an aerosol may be preferred, in particular in amounts of 70 micrograms, and particularly preferred in the amount of more than 250 or 1000 micrograms, or 5, 25 or 100 milligrams of oligonucleotide.

[0107] The combination of the adjuvant use of the oligonucleotides with adjuvant therapies such as carboplatin and pemetrexed, and the combination with radiotherapy may be preferred.

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