FIELD OF THE INVENTION

The present invention is in the field of a kit of parts comprising a radionuclide and a therapeutic compound, a medic ^¬ ament comprising said kit of parts, and a method of treating a disease by administering said kit of parts, in particular for treating a cancer or a metastase, wherein the therapeutic com ^¬ pound is released or activated.

BACKGROUND OF THE INVENTION

A radionuclide is an atom with an unstable nucleus, which is a nucleus characterized by excess energy available to be imparted either to a newly created radiation particle within the nucleus or to an atomic electron. The radionuclide, in this process, undergoes radioactive decay, and emits one or more of the following; photons, negatron, positron, or alpha particles, directly or indirectly. These particles constitute ionizing radiation. Radionuclides occur naturally, and can also be artificially produced.

Radionuclides are often referred to by chemists and physicists as radioactive isotopes or radioisotopes. Radioisotopes with suitable half-lives play an important part in a number of constructive technologies (for example, nuclear medicine) .

Radionuclides are used in two major ways: for their chemi- cal properties and as sources of radiation. Radionuclides of familiar elements such as carbon can serve as tracers because they are assumed to be chemically identical to the non-radioactive nuclides, so almost all chemical, biological, and ecological processes treat them in the same way.

Radioisotopes per se are also a method of treatment in hemopoietic forms of tumors; the success for treatment of solid tumors has been limited. More powerful gamma sources sterilize syringes and other medical equipment.

Cerenkov luminescence may be emitted when a charged parti- cle traverses a dielectric medium with a velocity greater than the phase velocity of light in the medium. The Cerenkov luminescence typically originates from β-radioisotopes . The light is then emitted on a cone around the particle direction, with a typical continuous spectrum. In tissue emitted light may be highly scattered and absorbed before reaching a surface, and the tissue's optical properties tend to favor the transmission of the red-infrared light, where Cerenkov emission is minimal.

Globally more than 8 million people may die from cancer every year. A majority thereof results from metastasized cancer cells, which are found extremely difficult to attack. At present metastasized cancer is treated primarily by chemotherapy. Since this therapy is systemic and tumor targeting is still far from optimal, chemotherapy may cause severe damage to (surrounding) healthy tissue, leading to rather adverse health effects including tissue damage, preliminary termina ^¬ tion of therapy programs and severe decrease in quality of life for the patient. One way to reduce side effects caused by undesired damage to healthy tissue is to design drug molecules or drug carriers that release the drug only at the tumor site, due to an internal or external trigger. Photodynamic therapy (PDT, light-induced activation of drugs using a sensitizer molecule) is one such example that has proven to be very ef ^¬ fective with limited adverse health effects. However, this therapy is limited to tumors of known location and, due to short penetration depth of light in tissue, to superficial tumors. Similarly, other external triggers can only treat localized solid tumors, while internal triggers rely on tumor dependent factors to release the drug, which often fail due to tumor microenvironment diversity.

Targeted therapy is a medical treatment typically used for cancer. Targeted therapy may be aimed at blocking growth of cancer cells or even destroy said cells. The term biologic therapy is sometimes synonymous with targeted therapy when used in the context of cancer therapy, which distinguishes therewith from chemotherapy. The two can be combined.

Another form of targeted therapy may involve use of nano- engineered enzymes to bind to a tumor cell thereby effectively eliminating it from the body. Targeted cancer therapies are considered more effective and less harmful to normal cells. Many targeted therapies may be considered as examples of immunotherapy developed by the field of cancer immunology.

Targeted therapies may relate to chemical entities that target or even preferentially target a protein or enzyme, such as one that carries a mutation or other genetic alteration that is specific to cancer cells and is hence not found in other host tissue. An example is a kinase inhibitor with ex ^¬ ceptional affinity for the oncofusion protein BCR-Abl.

There are targeted therapies for colorectal cancer, head and neck cancer, breast cancer, multiple myeloma, lymphoma, prostate cancer, melanoma and other cancers.

For drug delivery typically a vehicle or carrier is used. A carrier may relate to a liquid, such as water and oils, a dil ^¬ uent, typically water-based, such as a saline solution.

Photo-electrochemical processes may involve transforming light into other forms of energy. An example thereof is photo- sensitization, which relates to transferring energy of absorbed light. After absorption the energy may be transferred to certain reactants. In particular this process may be used when chemical reactions require light certain wavelengths that are not readily available. In an example mercury (Hg) absorbs radiation at 184.9 and 253.7 nm. It is a commonly used sensitizer. So when for instance mercury vapor is mixed with a chemical compound as ethylene, and the compound is irradiated therewith, a photodecomposition of ethylene to acetylene occurs by transfer of energy to the ethylene molecules. Cadmium, noble gases, zinc, benzophenone, and a large number of organic dyes may also be used as sensitizers. Photosensitizers are a key component of photodynamic therapy used to treat cancers.

Recently use of Cerenkov luminescence (CL) with nanotech- nology has been topic of a publication in Nature Nanotechnol- ogy by Shaffer et al . (7-2-2017), especially in view of imaging. Nanoparticles are described in combination with certain Cerenkov luminescence compounds for fluorescence imaging. Na- noparticles are considered best suited for use with CL given their high optical cross-sections compared with single small- molecule fluorophores . Examples of images obtained from experiments on mice are given. CL has also been subject of two patent applications having inventors in common. US2017007724 (Al) and WO2017019520 (Al) describe a two-component method, one component being a Cerenkov radiation sensitive component that may contain active payload, whereas the other component being a Cherenkov radiation emitting component. Therein the payload carrying component is the targeted component. The doc ^¬ uments are focused on illumination with ¹⁸ F. These method are found not to be very effective, for instance as a chance that the pay-load is released is too small. In addition only inac- tive compounds can be delivered, as a blocking agent is required. Also the radiation emitting compound has to be deliv ^¬ ered at or near (<imm) of the tumor, which is typically impossible or not feasible. In addition it is noted that the present inventors have found that the imaging compounds used are not stable under light conditions and in addition that these compounds do not dissolve very well and therefore can not be applied. A controlled release of payload is at least cumbersome. It is noted that for imaging Cerenkov nuclides have limited applicability in view of absorption by tissue. It suffers from all the drawbacks of optical imaging {reduced penetration depth and complex transport in tissue) , with additional limitations due to the broader emission spectrum and to the nonlinear correlation with the distribution of the primary source responsible of the Cerenkov emission.

Hartl (in J. Environmental Pathology, Toxicology and Oncology, Vol. 35, No. 2, 2016, p.185-192) recites yttrium ⁹⁰ being used to activate photodynamic therapy by generating Cerenkov radiation. Bauer (in M. Sc thesis, 2016, ProQuest LLC, Ann Arbor, Mi, 2016) recites exploration of Cerenkov luminescence activated photochemical internalization in cancer therapy.

Kamkaew (in ACS Applied Material & Interfaces, Vol. 8, No. 40, October 12 2016, p. 26630-26637) recites by Cerenkov Radiation induced photodynamic therapy using chlorin e6-loaded hollow mesoporous silica nanoparticles .

The above three documents suffer from (further) drawbacks, such as relatively high payloads need to be provided, such as a drug or medicament, and typically an injection into a tumour is used.

The present invention relates to a kit of parts, a medica- ment comprising said kit of parts, and a method of treating a disease by administering said kit of parts, which overcome one or more of the above disadvantages, without jeopardizing functionality and advantages. SUMMARY OF THE INVENTION

The present invention relates in a first aspect to a kit of parts according to claim 1. The above documents do not relate to two separate solution, each comprising a combination of various active compounds (e.g. radionuclide, targeting agent, photosensitizer, vehicle, and therapeutic compound) , which two solutions and compounds mutually interact. It is noted that especially the present vehicle and radionuclide, and also the photosensitizer and radionuclide, are in a sepa ^¬ rate solution, contrary to typical prior art. In addition the above documents at the best recite a few of the present active compounds, but not all. Some documents describe a radiation emitting component itself as a targeted component, and also important they do not describe Cerenkov radiation to trigger release of the payload; instead the payload is targeted to a cell or a group of cells. It is now found that significantly less payload, especially a drug or medicament can be used to be as effective as in the prior art. The payloads, such as a drug or medicament, are more effective, as release of the pay- load is controlled and limited to a desired location, and typically an injection, such as into a tumour, is not required. Typically an amount to payload can therefore be smaller compared to prior art administration routes. In addition as the delivery is in a controlled manner limited to the desired location, direct harmful effects of the payload to other areas of the body are virtually absent. In addition payload can be administered a few sequential times, wherein e.g. the β-parti- cle emitting radionuclide remains active over time; therewith a concentration of active compounds can be much lower as well. Also for the above prior art methods typically an injection into a tumour is used, as the radiation needs to be provided close to a tumour, which is not necessary with the present kit and method. The present kit thereto comprises a first solution, typically an aqueous solution, which is to be administered separately from a second solution, also typically an aqueous solution. The first solution comprises a radionuclide coupled to a targeting agent. The radionuclide is typically non-toxic to a human or animal body. The radionuclide-target- ing agent finds its way, after administration, through the body towards an intended cell receptor to which the targeting agent binds chemically. The radionuclide is capable of gener ^¬ ating Cerenkov radiation, i.e. photons, upon release of e.g. a β-particle. It is noted that the radiation of radionuclides it- self may also destroy or change the present vehicle, such as by cleaving a UV-cleavable group, such as an o-nitrobenzyl group and thioketals, such that the therapeutic compound is released; as such it may support the photosensitizer in its action. Specific radionuclides are found suitable, such as ¹³ N, ¹⁵ 0, ¹⁸ F, ⁴ Na, ^2S A1, ³¹ Si, ³² P, ³⁷ S, ³⁸ C1, ⁴² K, ⁴⁷ Ca, «Ca , ⁴⁴ Sc, ⁴⁷ Sc, ⁵¹ Ti, ⁵⁶ Mn, ⁶⁰ Co, ⁶² Cu, ⁶⁴ Cu, 67Cu, ^6S Ga, ⁶⁸ Ge, ⁷¹ Zn, ⁷⁰ Ga, ⁷² Ga, ⁷⁵ Ge, ⁷⁶ As, ⁸¹ Se, ⁸² Rb, ⁸³ Se, ⁸⁶ Rb, ⁸⁷ Kr, ⁸⁸ Rb, ⁸⁹ Sr, ⁸⁹ Zr, ⁹⁰ Y, ⁹⁵ Zr, "Mo, ¹⁰⁵ Ru, ¹¹⁰ Ag, ^min Pd, ^l Pd, ⁱⁿ Ag, ¹¹⁴ In, ¹²³ Sn, ^{1 4} In, ¹²⁵ Sn, ¹²⁷ Te, ^{1 6} I, ¹²⁸ I, ¹³¹ I, ¹³ Xe, ¹³⁵ Xe, ¹³⁹ Ba, ^{1 0} La, ¹⁴¹ La, ¹⁴¹ Ce, ¹⁴² Pr, ¹⁴⁹ Nd, ¹⁵¹ Nd, ¹⁵³ Sm, ¹⁵⁵ Sm, ¹⁵² Eu, ¹⁵ Eu, ¹⁵⁹ Gd, ^ieo Tb, ¹⁶⁵ Dy,

166 _Dyi 166 _Ho , "2 _Erf 1 7 _Ybi 177 _LUf 186 _Re , 187 _W/ 188 _{Re f} 191 _0s , 192 _ΐΓ

^1M Ir, ¹⁹⁷ Pt, ⁱ⁹⁸ Au, ²¹⁰ Bi, ²¹² Bi, ²¹³ Bi, ²³ Ra, ²²⁷ Th, and daughters of ²²⁵ Ac, preferably ¹³ N, * ⁵ 0, ³ P, ⁶⁰ Co, ⁶² Cu, ⁶⁸ Ga, ^S8 Ge, ⁸⁶ Rb, ⁸⁹ Sr, ⁸⁹ Zr, ⁹⁰ Y, ¹¹ In, ¹²⁴ In, ¹⁶⁶ Ho, ^$ Re, ¹⁸⁸ Re, ¹² Bi, ²¹³ Bi, ^{2 3} Ra, and daughters of ²²⁵ Ac. These radionuclides are found to give a good release of the therapeutic compounds (being present in the second solution), especially the preferred ones. Therewith only minute quantities of radionuclide are found to be required in view of an effective therapy. Such is a big ad- vantages as side-effects of therapeutic compounds, side-effects typically being present, are largely mitigated. It is noted that the radionuclide may be provided as a mixture of isotopes of the same chemical element; only the relevant isotopes are included in the above quantity of 10 ^~12 -10 ^~6 gr/1 (about 10- ¹⁴ -10 ^"8 mole/1) . The RN is coupled to a targeting agent, typically in a 1:1 ratio (mole/mole), though depending on the type of targeting agent other ratios are possible {e.g. 2 RN to 1 targeting agent molecule) . The targeting agent may therefore be present in an amount of 10- ¹ -10 ^~8 mole/1. The sec- ond solution comprises a relatively small amount of vehicle, typically 10 ^"4 -10 ¹ gr/1. The vehicle comprises 10" ⁵ -10° gr/gr of at least one therapeutic compound which is incorporated therein, such as enclosed, largely or fully surrounded by the vehicle, bound to the vehicle, forming part of the vehicle, such as a chemical entity thereof, and combinations thereof. A therapeutic compound molecule may be released, but also a rel ^¬ atively large number of molecules may be released at substan ^¬ tially the same time, preferably 2-100000 molecules, such as 100-10000 molecules. The vehicle further comprises lO^-lO ^-1 gr/gr of at least one photo-sensitizer , wherein the photo-sen- sitizer is incorporated in the vehicle or forms part of the vehicle, such as in the form of a chemical moiety. The at least one photo-sensitizer can be activated by the radionu- elide of the first solution, wherein the radionuclide typically emits a photon. When the vehicle is in the neighbourhood of the present radionuclide, which is (previously) attached to a cell by the targeting agent, a photon that originates from radionuclide as Cerenkov radiation can activate the photo-sen- sitizer. The photo-sensitizer initiates a structural change in the vehicle, which structural change can be a chemical change, a physical change, a partly or fully decomposition of the vehicle, and combinations thereof. By changing the structure of the vehicle the therapeutic compound is released at an in- tended location in the body and becomes effective at said location. Such is particularly suited for small sized tumours to be treated and metastases of tumours.

The present photosensitizer may be encapsulated into the present vehicle without a further need of modification, or may be conjugated to the present vehicle or carrier. A photosensitizer can be introduced to the carriers in various ways. In a first case, no modifications are necessary, block copolymer vesicle or micelle can be prepared using conventional methods in which a photosensitizer such as chlorin e6 can be incorpo- rated together with a chemotherapeutic drug. A photosensitizing molecule can also be conjugated at the end group of a block copolymer (e.g. for instance the FDA approved Pluronic F127) . Self-assemblies can be prepared with these conjugated polymers and when excited by light generate singlet oxygen de- stroying the nano-carrier . Photosensitizers considered are chlorin e6 and benzoporphyrin derivatives in view of their high quantum yield at excitation wavelengths in the UV spectrum (between 350 and 450 run) , matching the Cerenkov light spectrum. The block copolymer choice may depend on several factors. Block copolymers that are more likely to be accepted in clinical trials such as polyethylene oxide-polypropylene oxide-polyethylene oxide (i.e. Pluronic) , as well as polymers that are more prone to breaking upon singlet oxygen interaction such as polyethylene oxide-polystyrene (PEO-PS) are considered in particular. The exemplary photosensitizer molecule (e.g. chlorin e6) is linked to either the hydrophobic or the hydrophilic block and may require simple chemical modifications. For instance, chlorin e6 needs to be activated with ethyl dimethylaminopropyl carbondiamide in order to be attached to amino-modified polystyrene.

Likewise drugs may be conjugated to the present vehicles. For instance a hydroxyl group of an PEO block is converted to an amino group allowing the direct conjugation of the thioketal linkage. Drugs such as DOX can be added to the other side of the thioketal by small a modification of the drug, i.e. the addition of a carboxyl group.

The present kit of parts may be used in the manufacture of a medicament, such as a medicament for treatment of cancer, treatment of metastases, such as deeply located metastases and not-identified metastases. Likewise the present invention relates to a medicament comprising said kit of parts.

In a further aspect the present invention relates to a method of treating of a disease, in particular a cancer or me- tastase, comprising providing 0.1-5 ml/kg of the present second solution with the vehicle, administering 0.1-5 ml/kg of a first solution of the present invention with radionuclide coupled to a targeting agent for a tumour cell receptor, coupling the targeting agent to a tumour cell receptor, and activating the vehicle for release of the at least one therapeutic compound, preferably using timed sequential administration, such as with an idle time in between the provisions of 30 min-72 hours, such as 60 min-24 hours. The solutions are preferably administered intravenous, intratoneal, peritoneal, and sub-cutaneous .

Thereby the present invention provides a solution to one or more of the above mentioned problems.

Advantages of the present invention are detailed throughout the description. DETAILED DESCRIPTION OF THE INVENTION

The present invention relates in a first aspect to a kit of parts according to claim 1.

In an exemplary embodiment of the present kit the tar- geting agent may be able to bind to a cell receptor, such as a tumour cell receptor, such as a cell surface receptor, such as a prostate specific membrane antigen, such as Glu- NH-CO-NH-Lys- (Ahx) - ⁶⁸ Ga- (HBED-CC) . Exemplary receptors are ion channel linked receptor, such as cys-loop receptors, iono- tropic glutamate receptors and ATP-gated channels, an enzyme- linked receptor, such as an Erb receptor, such as ErbBl,

ErbB2, ErbB3, and ErbB4, a GDNF receptor, such as GFRal,

GFRa2 , GFRa3, and GFRa4, a NPR receptor, such as NPR1, NPR2, NPR3, and NPR4, a trk neurotrophin receptor, such as TrkA, TrkB, TrkC, and p75, and a toll-like receptor, such as TLR1, TLR2 , TLR3, TLR4, TLR5, TLR6, TLR7, TLR8 , TLR9, and TLR 10, and a G protein-coupled receptor, such as a rhodopsin-like receptor, such as chemokine (C-C motif) receptor 1, chemokine {C-C motif} receptor 2, chemokine (C-C motif) receptor 3, chemokine (C-C motif) receptor 4, chemokine (C-C motif) receptor 5, chemokine (C-C motif) receptor 6, chemokine (C-C motif) receptor 7, chemokine (C-C motif) receptor 8, chemokine (C-C motif) receptor 9, chemokine (C-C motif) receptor 10, chemokine (C-C motif) receptor-like 1, chemokine (C-C motif) recep- tor-like 2, chemokine (C motif) receptor 1, chemokine (C-X3-C motif) receptor 1, chemokine (C-X3-C motif) receptor 3, chemokine (C-X3-C motif) receptor 4, chemokine (C-X3-C motif) receptor 5, chemokine (C-X3-C motif) receptor 6, chemokine (C- X3-C motif) receptor 7, GPR137B, IL8R-alpha, IL8R-beta, andre- nomedullin receptor, duffy blood group chemokine receptor, G protein-coupled receptor 30, angiotensin II receptor type 1, angiotensin II receptor type 2, apelin receptor, bradykinin receptor Bl, bradykinin receptor B2, GPR15, GPR25, delta opioid receptor, kappa opioid receptor, mu opioid receptor, Noci- ceptin receptor, somatostatin receptor 1, somatostatin receptor 2, somatostatin receptor 3, somatostatin receptor 4, somatostatin receptor 5, neuropeptides B/W receptor 1, neuropeptides B/W receptor 2, galanin receptor 1, galanin receptor 2, galanin receptor 3, relaxin/insulin-like family peptide recep ^¬ tor 1, relaxin/insulin-like family peptide receptor 2, re ^¬ laxin/insulin-like family peptide receptor 3, relaxin/insulin- like family peptide receptor 4, KiSSl-derived peptide recep- tor, melanin-concentrating hormone receptor 1, urotensin-II receptor, cholecystokinin A receptor, cholecystokinin B receptor, neuropeptide FF receptor 1, neuropeptide FF receptor 2, hypocretin receptor 1, hypocretin receptor 2, arginine vaso ^¬ pressin receptor 1A, arginine vasopressin receptor IB, argi- nine vasopressin receptor 2, gonadotrophin releasing hormone receptor, pyroglutamylated RFamide peptide receptor, bombesin- like receptor 3, neuromedin B receptor, gastrin-releasing pep ^¬ tide receptor, endothelin receptor type A, endothelin receptor type B, GPR37, neuromedin U receptor 1, neuromedin U receptor 2, neurotensin receptor 1, neurotensin receptor 2, thyrotro- pin-releasing hormone receptor, growth hormone secretagogue receptor, Motilin receptor, C3a receptor, C5a receptor, chemo- kine-like receptor 1, formyl peptide receptor 1, formyl peptide receptor-like 1, formyl peptide receptor-like 2, MAS1, MAS1L, GPR1, GPR32, tachykinin receptor 1, tachykinin receptor 2, tachykinin receptor 3, neuropeptide Y receptor Yl, neuropeptide Y receptor Y2, pancreatic polypeptide receptor 1, neuropeptide Y receptor Y5 , prolactin-releasing peptide receptor, prokineticin receptor 1, prokineticin receptor 2, GPR19,

GPR50, FSH-recepto , luteinizing horitvone/choriogonadotropin receptor, thyrotropin receptor, melanocortin 1 receptor, mela- nocortin 3 receptor, melanocortin 4 receptor, melanocortin 5 receptor and a ACTH receptor. Preferably the receptor is a receptor that, compared to non-tumour cells, is expressed more abundant at a tumour cell. Therewith very specific and effective binding of the targeting agent and radionuclide coupled thereto is achieved.

In an exemplary embodiment of the present kit the targeting agent may be selected from a peptide or protein that is able to bind to a cell receptor. Examples thereof are epidermal growth factor, glial cell-derived neurotrophic factor, natriuretic peptide, a neurotropin, such as nerve growth factor, brain-derived neurotrophic factor, neurotrophin-3, and neuro- trophin-4, a microbial peptide or protein ligand, a chemokine, such as a chemokine, such as a homeostatic chemokine, such as CCL14, CCL19, CCL20, CCL21, CCL25, CCL27, CXCL12 and CXCL13 , and an inflammatory chemokine, such as CXCL8, CCL2, CCL3, CCL4, CCL5 , CCL11 and CXCL10, andrenomedullin, an angiotensin, such as angiotensin I, angiotensin II, angiotensin III and angiotensin IV, apelin, bradykinin, an opioid peptide, such as an enkephalin, an endophin and an dynorpin, nociceptin, a neuropeptide, such as galanin, somatostatin, neuropeptide Y, cholecystokinin, vasoactive intestinal peptide, a tachykinin peptide, such as neurokinin A and neurokinin K, neuropeptide gamma, and substance P, neurotensin, glucagon-like peptide-1, hypocretin and thyrotropin-releasing hormone, relaxin, insulin, kisspeptin, melanin-concentrating hormone, urotensin-II , vasopressin, gonadotrophin, bombesin, neuromendin B, neu- romendin U, gastrin, endothelin, growth hormone, a growth hormone secretagogue, such as ghrelin, pralmorelin, GHRP-6, ex- amorelin, ipamorelin, ibutamoren, growth hormone-releasing hormone, CJC-1295, sermorelin, and tesamorelin, motilin, C3a, C5a, an N-formyl peptide, chemerin, pancreatic polypeptide, prolactin-releasing peptide, prokineticin, follicle-stimulating hormone, luteinizing hormone, choriogonadotropin, thyrotropin and a melanocortin, such as adrenocorticotropic hormone and a melanocyte-stimulating hormone.

In an exemplary embodiment of the present kit the radionuclide may be selected from ¹³ N, ¹⁵ 0, ³² P, ⁶² Cu, ⁶⁸ Ga, ⁶⁸ Ge, ⁸⁶ Rb, ⁸⁹ Sr, 9° _Yf n«in, ¹⁶⁶ Ho, ^18S Re, ²¹ Bi, ²¹³ Bi, ²²³ Ra, daughters of ²²⁵ Ac, and ²²⁷ Th, preferably ¹⁵ 0, ² P, ^S2 Cu, ⁶⁸ Ga, ⁶⁸ Ge, ⁸⁶ Rb, ^ss Sr, ⁹⁰ Y, ¹¹⁴ In, !66Ho, ^18S Re, ²¹² Bi, 223 _Ra , _and 22? _{Thi more} preferably ¹⁵ 0, 62 _CU/ 68 _Gaf 68 _G e _f 86 _Rb/ 9Cy _f 114 _Iri/ 166 _Ηθ/ 188 _Re , _and 212 _Bi/ even more preferably ^S2 Cu, ⁶⁸ Ga, ⁶⁸ Ge, ⁸⁶ Rb, ⁹⁰ Y, ⁴ In, ¹⁶⁶ Ho, and ¹⁸⁸ Re, such as ⁶⁸ Ga and ⁸² Rb. These radionuclides activate the present photo-sensitizer well and therewith establish an effective release of the present therapeutic compound.

In an exemplary embodiment of the present kit the vehicle may be selected from nano-carriers and micro-carriers, such as microspheres, liposomes, pol merosomes, micelles, silica, hybrid particles, dendrimers, carbon nanoparticles, such as fullerenes, inorganic particles, such as hydrophobic-hydro- philic block-copolymer micelles, such as azobenzene comprising block-copolymers, such as a copolymer of poly {tert-butyl acry- late-coacrylic acid) and polymethacrylate bearing azobenzene, poly (ethylene oxide) (PEO) as the hydrophilic block and a polymethacrylate bearing spiropyran moieties or polybutadiene as the hydrophobic block, and combinations thereof. Further examples are polyethylene oxide-polylactic acid (PEO-PLA) , polyethylene oxide-polystyrene (PEO-PS) , and pol { 1-pyrenylme- thyl methacrylate) { PPy A) as hydrophilic block. The present vehicle may be amphiphilic. Hydrophilic blocks, such as PEO's, may have a molecular weight of 1000-10000 g/mole, such as 2500-6000, e.g. 4000-5000, whereas the hydrophobic blocks, such as PB, may have a molecular weight of 1000-10000 g/mole, such as 1500-6000, e.g. 1800-4000. The nano-carriers typically have a cross-section of 10-500 nm, whereas the micro-carriers typically have a cross-section of 0.5-100 um. They may relates to organic molecules, such as liposomes, polymerosomes, etc. and be of inorganic type, such as silica, and combinations thereof. The vehicles can be loaded with a therapeutic compound by using a chemical technique, such as absorption of the therapeutic compound being present in a liquid, and by physical techniques, such as deposition of the therapeutic compound on the vehicle.

As mentioned previously an important part is to ensure that the drug is released primarily at the tumour site and not affecting healthy tissue. This means that e.g. a nano-carrier may need to be very stable, having little drug loss before being triggered. Also, the radionuclide should be brought to the tumour using a targeting vector which preferably does not accumulate at organs of the MPS (Mononuclear phagocyte system) typically involved in nano-carriers' clearance. Nano-carriers rely on the so-called Enhanced Permeation and Retention (EPR) effect to accumulate at the tumour site, also called passive targeting. The EPR effect is known to occur for certain malignancies when the tumour reaches a certain size and requires more nutrients to grow. New blood vessels are then created, which are often defective, i.e. their endothelial cells are not tightly aligned allowing the escape of molecules. Since tumours lack lymphatic drainage, large entities such as nano- carriers can accumulate at the diseased site provided that they circulate long enough in the blood. There are several pa ^¬ rameters that are recognized to play a role in blood circulation of nanoparticles, such as size, shape, flexibility, surface properties etc. Large particles (> 200 rati) are rapidly cleared by the MPS, while small ones (< 10 nm) rapidly undergo renal clearance, limiting those suitable for the EPR effect to sizes typically ranging between 10 and 200 nm, e.g. 20-100 nm. Surface modification with polyethylene glycol (PEG, also called polyethylene oxide) may be carried out to decrease in- teraction with macrophages and a PEG molecular weight above

2000 g/mol is considered sufficient to increase blood circula ^¬ tion half-lives. Inventors prepared micelles or polymer vesicles (polymersomes ) and to a lesser extent rod or worm-like micelles, since the benefit of larger aspect ratios in polymer systems has not been completely convincing. Low molecular weight targeting vectors commonly used in radionuclide therapy such as various peptides, antibody fragments and small molecules, rapidly reach their target while a remainder is cleared within a short time by the kidneys and subsequently excreted. This means that if the radionuclide is bound to a low molecular weight targeting vector and administered after the nano- carrier has been cleared from the blood, the nano-carrier and radionuclide will mainly encounter each other at the tumour.

In an exemplary embodiment of the present kit the vehi- cle may comprise a layer, which layer is adapted to at least partly disintegrate or undergo a structural change upon direct or indirect activation by the photo-sensitizer . The change may relate to a transition from e.g. a hydrophilic to a hydrophobic state, to an opening of the layer, to disruption of chemi- cal bonds, resulting in the release of the therapeutic compound. The layer may have a thickness of 10-10000 nm.

In an exemplary embodiment of the present kit the photo-sensitizer may comprise at least one of an aromatic ring, such as 1-5 aromatic rings, a conjugated double bond, a conjugated triple bond, a C=N bond, a isomeric changeable C=C cis or trans bond, such as violanthrone, isoviolanthrone, fluorescein, rubrene, 9, 10-diphenylanthracene, tetracene, 13,13'- dibenzantronile, acrylcarbonylmethyl , nitrtoaryl, coumarin, arylmethyl, and levulinic acid. The present photo-sensitizer (PS) preferably absorbs light in the wavelength region of 180- 500 nm, such as 200-360 ran, it provides a high singlet oxygen quantum yield, the excited triplet state has a long lifetime to allow for sufficient interaction between ground state oxy- gen and excited PS, it has a high resistance to photo-bleach- . ing, it has a high resistance to oxidation by ¹ 02, the energy of the excited triplet state (difference between the Ti and the So state) is ≥ 94 kj/mole, which is the energy required to con ^¬ vert ground state oxygen to ¹ 02, and is non-toxic, metabolizable and inert. In addition at neutral or slightly acidic/basic conditions (pH 5-9) the PSs preferably do not aggregate, such as in concentrations of 10-500 mM. Good examples are

phenalenone and chlorin e6.

In an exemplary embodiment of the present kit the photo-sensitizer may produce a singlet oxygen, is cleavable by a UV-photon, or initiates a cis-trans transition.

In an exemplary embodiment of the present kit the at least one therapeutic compound may be incorporated in a nano- carrier, such as a nano-carrier or micro-carrier identified above.

In an exemplary embodiment of the present kit the therapeutic compound may be selected from at least one of medicaments, drugs, chemotherapeutics , genes, metabolic active compound (TBC) , and antibiotics, or a derivative thereof, or an analogue thereof. Also pro-drugs may be considered. In an exemplary embodiment of the present kit the drug is selected from cancer drugs, and cardio drugs. Examples of these drugs are cancer drugs, such as Abiraterone Acetate, Abitrexate or Methotrexate, Abraxane, Adcetris or Brentuximab Vedotin, Ado- Trastuzumab Emtansine, Adriamycin or Doxorubicin Hydrochloride, Afatinib Dimaleate, Afinitor or Everolimus, Akynzeo or Netupitant and Palonosetron Hydrochloride, Aldara or

Imiquimod, Aldesleukin, Alecensa or Alectinib, Alectinib,

Alemtuzumab, Alkeran for Injection or Melphalan Hydrochloride, Alkeran Tablets or Melphalan, Alimta or Pemetrexed Disodium,

Aloxi or Palonosetron Hydrochloride, Ambochlorin or Chlorambucil, Amboclorin or Chlorambucil, Amifostine, Aminolevulinic Acid, Anastrozole, Aprepitant, Aredia or Pamidronate Disodium, Arimidex or Anastrozole, Aromasin or Exemestane, Arranon or Nelarabine, Arsenic Trioxide, Arzerra or Ofatumumab, Asparagi ^¬ nase Erwinia chrysanthemi, Atezolizumab, Avastin or Bevaci- zumab, Avelumab, Axitinib, Azacitidine, Bavencio or Avelumab, BEACOPP, Becenum or Carmustine, Beleodaq or Belinostat, Beli- nostat, Bendamustine Hydrochloride , BEP, Bevacizumab, Bexaro- tene, Bexxar or Tositumomab and Iodine I 131 Tositumomab, Bi- calutamide, BiCNU or Carmustine, Bleomycin, Blinatumomab, Blincyto or Blinatumomab, Bortezomib, Bosulif, Bosutinib, Brentuximab Vedotin, BuMel, Busulfan, Busulfex, Cabazitaxel, Cabometyx, Cabozantinib-S-Malate, Campath or Alemtuzumab,

Camptosar or Irinotecan Hydrochloride, Capecitabine, Carac or Fluorouracil— opical, Carboplatin, CARBOPLA IN-TAXOL, Carfil- zomib, Carmubris, Carmustine, Casodex or Bicalutamide,

Ceritinib, Cerubidine or Daunorubicin Hydrochloride, Cervarix or Recombinant HPV Bivalent Vaccine, Cetuximab, Chlorambucil, CHLORAMBUCIL-PREDNISONE, Cisplatin, Cladribine, Clafen or Cyclophosphamide, Clofarabine, Clofarex, Clolar, Cobimetinib, Cometriq or Cabozantinib-S-Malate, COPDAC, COPP, COPP-ABV, Cosmegen or Dactinomycin, Cotellic or Cobimetinib, Crizotinib, CVP, Cyclophosphamide, Cyfos or Ifosfamide, Cyramza or Ramu- cirumab, Cytarabine, Cytarabine Liposome, Cytosar-U or Cytara- bine, Cytoxan or Cyclophosphamide, Dabrafenib, Dacarbazine, Dacogen or Decitabine, Dactinomycin, Daratumumab, Darzalex, Dasatinib, Daunorubicin Hydrochloride, Decitabine, Defibrotide Sodium, Defitelio, Degarelix, Denileukin Diftitox, Denosumab, DepoCyt or Cytarabine Liposome, Dexamethasone, Dexrazoxane Hydrochloride, Dinutuximab, Docetaxel, Doxil or Doxorubicin Hydrochloride Liposome, Doxorubicin Hydrochloride, Doxorubicin Hydrochloride Liposome, DTIC-Dome or Dacarbazine, Efudex or Fluorouracil—Topical, Elitek or Rasburicase, Ellence or Epi- rubicin Hydrochloride, Elotuzumab, Eloxatin or Oxaliplatin, Eltrombopag Olamine, Emend or Aprepitant, Empliciti or Elotuzumab, Enzalutamide, Epirubicin Hydrochloride, EPOCH,

Erbitux or Cetuximab, Eribulin Mesylate, Erivedge or Vismo- degib, Erlotinib Hydrochloride, Erwinaze or Asparaginase Erwinia chrysanthemi, Ethyol or Amifostine, Etopophos or Etopo- side Phosphate, Etoposide, Etoposide Phosphate, Evacet or Doxorubicin Hydrochloride Liposome, Everolimus, Evista or Raloxi- fene Hydrochloride, Evomela or Melphalan Hydrochloride, Ex- emestane, 5-FU or Fluorouracil Injection, 5-FU or Fluoroura- cil--Topical, Fareston or Toremifene, Farydak or Panobinostat, Faslodex or Fulvestrant, Femara or Letrozole, Filgrastim, Fludara or Fludarabine Phosphate, Fludarabine Phosphate,

Fluoroplex, Fluorouracil Injection, Fluorouracil—Topical , Flutamide, Folex, Folex PFS, FOLFIRI, FOLFIRI-BEVACIZUMAB, FOLFIRI-CETUXIMAB, FOLFIRINOX, FOLFOX, Folotyn or Pralatrex- ate, FU-LV, Fulvestrant, Gardasil or Recombinant HPV Quadriva- lent Vaccine, Gardasil 9 or Recombinant HPV Nonavalent Vac ^¬ cine, Gazyva or Obinutuzumab, Gefitinib, Gemcitabine Hydro ^¬ chloride, GEMCITABINE-CISPLATIN, GEMCITABINE-OXALI PLA IN, Gemtuzumab Ozogamicin, Gemzar or Gemcitabine Hydrochloride, Gilotrif or Afatinib Dimaleate, Gleevec or Imatinib Mesylate, Gliadel or Carmustine Implant, Gliadel wafer or Carmustine Implant, Glucarpidase, Goserelin Acetate, Halaven or Eribulin Mesylate, Hemangeol or Propranolol Hydrochloride, Herceptin or Trastuzurnab, HPV Bivalent Vaccine, Recombinant HPV Nonavalent Vaccine, Recombinant HPV Quadrivalent Vaccine, Recombinant Hycamtin or Topotecan Hydrochloride, Hydrea, Hydroxyurea, Hy- per-CVAD, Ibrance or Palbociclib, Ibritumomab Tiuxetan Ibru- tinib, Iclusig or Ponatinib Hydrochloride, Idamycin, Idarubi- cin Hydrochloride, Idelalisib, Ifex, Ifosfamide, Ifosfamidum, IL-2 or Aldesleukin, Imatinib Mesylate, Imbruvica or Ibru- tinib, Imiquimod, Imlygic or Talimogene Laherparepvec, Inlyta or Axitinib, Interferon Alfa-2b, Recombinant Interleukin-2 or Aldesleukin, Intron A or Recombinant Interferon Alfa-2b, Iodine I 131 Tositumomab and Tositumomab, Ipilimumab, Iressa or Gefitinib, Irinotecan Hydrochloride, Irinotecan Hydrochloride Liposome, Istodax or Romidepsin, Ixabepilone, Ixazomib Citrate, Ixempra or Ixabepilone, Jakafi or Ruxolitinib Phosphate, Jevtana or Cabazitaxel, Kadcyla or Ado-Trastuzumab Emtansine, Keoxifene or Raloxifene Hydrochloride, Kepivance or Palifer- min, Keytruda or Pembrolizumab, Kisqali or Ribociclib, ypro- lis or Carfilzomib, Lanreotide Acetate, Lapatinib Ditosylate, Lartruvo or Olaratumab, Lenalidomide , Lenvatinib Mesylate, Lenvima or Lenvatinib Mesylate, Letrozole, Leucovorin Calcium, Leukeran or Chlorambucil, Leuprolide Acetate, Leustatin or Cladribine, Levulan or Aminolevulinic Acid, Linfolizin or Chlorambucil, LipoDox or Doxorubicin Hydrochloride Liposome, Lomustine, Lonsurf or Trifluridine and Tipiracil Hydrochlo ^¬ ride, Lupron or Leuprolide Acetate, Lupron Depot or Leuprolide Acetate, Lupron Depot-Ped or Leuprolide Acetate, Lynparza or Olaparib, Marqibo or Vincristine Sulfate Liposome, Matulane or Procarbazine Hydrochloride, Mechlorethamine Hydrochloride, Megestrol Acetate, Mekinist or Trametinib, Melphalan, Melpha- lan Hydrochloride, Mercaptopurine, Mesna, Mesnex, Methazolas- tone or Temozolomide, Methotrexate, Methotrexate LPF, Methyl- naltrexone Bromide, Mexate, Mexate-AQ, Mitomycin C, Mitoxan- trone Hydrochloride, Mitozytrex, Mozobil or Plerixafor, Mus- targen or Mechlorethamine Hydrochloride, Mutamycin or Mitomycin C, Myleran or Busulfan, Mylosar or Azacitidine, Mylotarg or Gemtuzumab Ozogamicin, Navelbine or Vinorelbine Tartrate, Necitumumab, Nelarabine, Neosar or Cyclophosphamide, Netupi- tant and Palonosetron Hydrochloride, Neulasta or Pegfil- grastim, Neupogen or Filgrastim, Nexavar or Sorafenib Tosyl- ate, Nilandron, Nilotinib, Nilutamide, Niniaro or Ixazomib Citrate, Nivolumab, Nolvadex or Tamoxifen Citrate, Nplate or Romiplostim, Obinutuzumab, Odomzo or Sonidegib, Ofatumumab, Olaparib, Olaratumab, Omacetaxine Mepesuccinate, Oncaspar or Pegaspargase, Ondansetron Hydrochloride, Onivyde or Irinotecan Hydrochloride Liposome, Ontak or Denileukin Diftitox, Opdivo or Nivolumab, Osimertinib, Oxaliplatin, Paclitaxel, Palbo- ciclib, Palifermin, Palonosetron Hydrochloride, Netupitant, Pamidronate Disodium, Panitumumab, Panobinostat , Paraplat or Carboplatin, Pazopanib Hydrochloride, PCV, PEB, Pegaspargase Pegfilgrastim, Peginterferon Alfa-2b, PEG-Intron, Pembroli- zumab, Pemetrexed Disodium, Perj eta, Pertuzumab, Platinol or Cisplatin, Platinol-AQ, Plerixafor, Pcmalidomide, Pomalyst,

Ponatinib Hydrochloride, Portrazza or Necitumumab, Pralatrex- ate, Prednisone, Procarbazine Hydrochloride, Proleukin or Aldesleukin, Prolia or Denosumab, Promacta or Eltrombopag Olamine, Propranolol Hydrochloride, Provenge or Sipuleucel-T, Purinethol or Mercaptopurine, Purixan, Radium 223 Dichloride, Raloxifene Hydrochloride, Ramucirumab, Rasburicase, R-CHOP, R- CVP, Recombinant drugs, Recombinant Human Papillomavirus or HPV, Bivalent Vaccine, Recombinant Human Papillomavirus or HPV, Nonavalent Vaccine, Recombinant Human Papillomavirus or HPV, Quadrivalent Vaccine, Recombinant Interferon Alfa-2b, Regorafenib, Relistor or Methylnaltrexone Bromide, R-EPOCH, resveratrol, Revlimid or Lenalidomide, Rheumatrex or Metho ^¬ trexate, Ribociclib, R-ICE, Rituxan or Rituximab, Rituximab, Rolapitant Hydrochloride, Romidepsin, Romiplostim, Rubidomycin or Daunorubicin Hydrochloride, Rubraca, Rucaparib Camsylate, Ruxolitinib Phosphate, Sclerosol Intrapleural Aerosol, Siltux- imab, Sipuleucel-T, Somatuline Depot or Lanreotide Acetate, Sonidegib, Sorafenib Tosylate, Sprycel or Dasatinib, STANFORD V, Stivarga or Regorafenib, Sunitinib Malate, Sutent or

Sunitinib Malate, Sylatron or Peginterferon Alfa-2b, Sylvant or Siltuximab, Synribo or Omacetaxine Mepesuccinate, Tabloid or Thioguanine, Tafinlar or Dabrafenib, Tagrisso or Osimer- tinib, Talc, Talimogene Laherparepvec, Tamoxifen Citrate, Tar- abine PFS or Cytarabine, Tarceva or Erlotinib Hydrochloride, Targretin or Bexarotene, Tasigna or Nilotinib, Taxol or

Paclitaxel, Taxotere or Docetaxel, Tecentriq or Atezolizumab, Temodar or Temozolomide, Temozolomide, Temsirolimus, Thalidomide, Thalomid or Thalidomide, Thioguanine, Thiotepa, Tolak or Fluorouracil--Topical, Topotecan Hydrochloride, Toremifene, Torisel or Temsirolimus, Tositumomab and Iodine I 131 Tosi- tumomab, Totect or Dexrazoxane Hydrochloride, Trabectedin, Trametinib, Trastuzumab, Treanda or Bendamustine Hydrochloride, Trifluridine and Tipiracil Hydrochloride, Trisenox or Arsenic Trioxide, Tykerb or Lapatinib Ditosylate, Unituxin or Dinutuximab, Uridine Triacetate, Vandetanib, Varubi or Rolapitant Hydrochloride, Vectibix or Panitumumab, VelP, Velban or Vinblastine Sulfate, Velcade or Bortezomib, Velsar or Vinblastine Sulfate, Vemurafenib, Venclexta or Venetoclax, Veneto- clax, Viadur or Leuprolide Acetate, Vidaza or Azacitidine,

Vinblastine Sulfate, Vincasar PFS or Vincristine Sulfate, Vincristine Sulfate, Vincristine Sulfate Liposome, Vinorelbine Tartrate, Vismodegib, Vistogard or Uridine Triacetate, Vorax- aze or Glucarpidase , Vorinostat, Votrient or Pazopanib Hydro- chloride, Wellcovorin or Leucovorin Calcium, Xalkori or Crizo- tinib, Xeloda or Capecitabine, XELIRI, XELOX, Xgeva or Deno- sumab, Xofigo or Radium 223 Dichloride, Xtandi or Enzalutam- ide, Yervoy or Ipilimumab, Yondelis or Trabectedin, Zaltrap or Ziv-Aflibercept, Zarxio or Filgrastim, Zelboraf or Vemuraf- enib, Zevalin or Ibritumomab Tiuxetan, Zinecard or Dexrazoxane Hydrochloride, Ziv-Aflibercept , Zofran or Ondansetron Hydrochloride, Zoladex or Goserelin Acetate, Zoledronic Acid,

Zolinza or Vorinostat, Zometa or Zoledronic Acid, Zydelig or Idelalisib, Zykadia or Ceritinib, Zytiga or Abiraterone Acetate, in particular docetaxel, doxorubicin, paclitaxel, cis- platin, capox, carboplatin, Carfilzomib, Rucaparib, and Cam- sylate, cardio drugs, such as agents for hypertensive emergen ^¬ cies, for pulmonary hypertension, aldosterone receptor antagonists, angiotensin converting enzyme inhibitors, angiotensin receptor blockers, angiotensin receptor blockers, neprilysin inhibitors, for antiadrenergy, for centrally acting antiadren- ergy, for peripherally acting antianginal, for antiarrhyth ^¬ mics, such as so-called group I antiarrhythmics, group II antiarrhythmics, group III antiarrhythmics, group IV antiarrhythmics, and group V antiarrhythmics, for anticholinergic chronotropy, for antihypertensive combinations, such as ACE inhibitors and calcium channel blocking, ACE inhibitors with thiazides, angiotensin II inhibitors and calcium channel blockers, angiotensin II inhibitors and thiazides, for antiadrenergy or central, and thiazides, for antiadrenergic or peripheral, and thiazides, beta blockers and thiazides, for potassium sparing diuretics and thiazides, for beta-adrenergy blocking, such as cardio selective beta blockers, and non-car- dio selective beta blockers, for calcium channel blocking, catechol amines, diuretics, such as carbonic anhydrase inhibitors, loop diuretics, miscellaneous diuretics, potassium-sparing diuretics, thiazide diuretics, for inotropy, peripheral vasodilators, renin inhibitors, for sclerosy, vasodilators, vasopressin antagonists, vasopressors, and antibiotics, such as aminoglycosides, beta lactams, such as amoxicillin, car- bapenems, penicillins, cephalosporins, kanamycins, thienarrty- cin, doxycycline, cephalexin, ciprofloxacin, clindamycin, metronidazole, azithromycin, sulfamethoxazole, trimethoprim, amoxicillin, clavulanate, levofloxacin, fosfomycins, mycines, such as amikacin, arbekacin, azithtromycin, butirosin, clarithromycin, dibekacin, erythromycin, kanamycin, Kanamycin A, Kanamycin B, Kanamycin C, Kanamycin D, Kanamycin X, 3"-Deam- ino-3"-hydroxykanamycin B, 3"-Deamino-3"-hydroxykanamycin C, 3"-Deamino-3"-hydroxykanamycin X, lividomycin, Nebramycin 5', neomycin, neomycin B, neomycin C, neomycin E, ribostamycin, streptomycin, tobramycin, vancomycin, and micines, such as dexoystreptamine, geneticin, gentamicin, gentamicin A2, gen- tamicin Ci, gentamicin C2, gentamicin C18, isepamicin, netilmicin, and sisortiicinazabicyclo [ 3.2.0 ] hept-2-ene-2-carbox- ylic acids, such as 7-oxo-l-azabicyclo [3.2.0] hept-2-ene-2-car- boxylic acids, such as thienamycin ( ( 5R, 6S) -3- [ (2-Ami- noethyl) thio] -6- [ ( 1R) -1-hydroxyethyl ] -7-oxo-l-azabicyclo [ 3.2.0 ] hept-2-ene-2-carboxylic acid), imipenem (5R,6S}-6- [ (1R) -1-hydroxyethyl] -3- ( { 2- [ ( iminomethyl ) amino] ethyl}thio) -7- oxo-l-azabicyclo [3.2.0] hept-2-ene-2-carboxylic acid, meropenem 4R, 5S, 6S) -3- ( ( (3S, 5S) -5- (Dimethylcarbamoyl) pyrrolidin-3- yl) thio) -6- ( (R) -1-hydroxyethyl) -4-methyl-7-oxo-l-azabicy- clo [3 - 2.0] hept-2-ene-2-carboxylic acid, ertapenem (4R, 5S,6S)-

3- [ (3S, 5S) -5- [ { 3-carboxyphenyl ) carbamoyl] pyrrolidin-3-yl] sul- fanyl-6- (1-hydroxyethyl) -4-methyl-7-oxo-l-azabicy- clo [3.2.0] hept-2-ene-2-carboxylic acid, doripenem (4R,5S,6S)- 6- (1-Hydroxyethyl) -4-methyl-7-oxo-3- ( ( (5S) -5- ( (sul- famoylamino) methyl) pyrrolidin-3-yl ) thio) -1-azabicyclo [3.2.0] hept-2-ene-2-carboxylic acid, panipenem/betamipron ( 5R, 6S ) -3- { [ (3S) -l-ethanimidoylpyrrolidin-3-yl] sulfanyl } - 6- [ (1R) -1-hydroxyethyl] -7-oxo-l-azabicyclo [3- 2.0] hept-2-ene-2- carboxylic acid, biapenem (4R, 5S, 6S) -3- (6, 7-dihydro-5H- pyra- zolo [1, 2-a] [1, 2, 4] triazol-8- ium-6-ylsulfanyl) - 6- { 1-hydroxyethyl)- 4-methyl-7-oxo-l-azabicyclo [3.2.0] hept-2- ene-2-car- boxylate, razupenem (4R, 5S, 6S) -6- ( (R) -1-hydroxyethyl) -4-me- thyl-3- ( (4- ( (S) -5-methyl-2, 5-dihydro-lH-pyrrol-3-yl) thiazol-2- yl) thio) -7-oxo-l-azabicyclo [3.2.0] hept-2-ene-2-carboxylic acid, tebipenem (4R, 5S, 6S) - ( Pivaloyloxy) methyl 3- ( ( 1- ( 4 , 5-di- hydrothiazol-2-yl) azetidin-3-yl ) thio) -6- ( (R) -1-hydroxyethyl) -

4-methyl-7-oxo-l-azabicyclo [3.2.0] hept-2-ene-2-carboxylate, lenapenem, tomopenem ( (4R, 5S, 6S) -3- [ (3S, 5S) -5- [ (3S) -3- [ [2- (di- aminomethylideneamino) acetyl] amino] pyrrolidine-l-carbonyl] -1- methylpyrrolidin-3-yl] sulfanyl-6- [ ( 1R) -1-hydroxyethyl] -4-me- thyl-7-oxo-l-azabicyclo [3.2.0] hept-2-ene-2-carboxylic acid) , or a derivative thereof, or an analogue thereof. Hence a large variety of therapeutic compounds can be administered and se ^¬ lected for a given purpose.

In an exemplary embodiment of the present kit the radi ^¬ onuclide may produce photons with a maximum intensity at a wavelength of 180-500 nm, e.g. 200-360 nm. It has been found that especially these photons are effective in terms of structurally changing the present vehicle and as a result release of the therapeutic compound thereof.

In an exemplary embodiment of the present kit the radi- onuclide may have a half life time of 5 min-28 days. The ac ^¬ tivity of the radionuclide is preferably not too low and also the activity is maintained over a prolonged period of time. For many applications the half life time is not very critical; for the present invention the half life time is limited in view of the administration of the therapeutic compound, which is released only upon (indirect) generation of photons by the present radionuclide. In this respect especially ¹³ N, ³² P, ⁶² Cu, ^6S Ga, ^8S Rb, ⁸⁹ Sr, ⁹⁰ Y, ^16S Ho, ¹⁸⁸ Re, ²¹² Bi, ²¹³ Bi, ²²³ Ra, daughters of ²²⁵ Ac, and ²²⁷ Th are considered, preferably ³² P, ⁶⁸ Ga, ⁹⁰ Y, ¹⁶⁵ Ho, ^18S Re, ²¹² Bi, ²¹³ Bi, ²²³ Ra, and daughters of ²²⁵ Ac, more preferably ^S8 Ga, ^9C Y, ¹⁵⁶ Ho, ^1S8 Re, and ²¹² Bi, even more preferably ⁶⁸ Ga, ⁹⁰ Y,

166 _HOf 188 _{Re/ such as} 68 _Ga _

In an exemplary embodiment of the present kit the radionuclide may be conjugated to the targeting agent, such as to a peptide. Therewith a stable bond is provided which is maintained over the life time of the radionuclide.

In an exemplary embodiment of the present kit the radionuclide dose may be 0.1-200 Gy (J/kg = m ² /s ² ) , preferably 0.2- 150 Gy, more preferably 1-100 Gy, even more preferably 5-80 Gy, such as 10-50 Gy or 20-40 Gy. Therewith both large doses

{e.g. > 80 Gy, such as > 120 Gy) and small doses (e.g. < 1 Gy, such as < 0.5 Gy) can be provided; the present kit of parts is very versatile in this respect.

The invention is further detailed by the accompanying figures and examples, which are exemplary and explanatory of nature and are not limiting the scope of the invention. To the person skilled in the art it may be clear that many variants, being obvious or not, may be conceivable falling within the scope of protection, defined by the present claims .

SUMMARY OF THE FIGURES

Figure 1 shows schematics of the present invention.

Figure 2 shows Ce6 retention in PB-PEO (Mn^l800-4000 ) micelles upon γ-irradiation from a (GC200) ⁵⁰ Co source. In this case Ce6 functions as the singlet oxygen source as well as a dye to follow its release from the micelles.

DETAILED DESCRIPTION OF THE FIGURES

Figure 1 shows schematics of the present invention. Fig- ure 1 Cerenkov triggered drug (D) release at the tumour site mediated by a (worked open) nano-carrier (NC) (second solu ^¬ tion) responding to Cerenkov radiation (first solution) by activation of the photosensitizer (PS) . Fig. 1A shows the nano-carrier accumulated at the tumour and fig. IB the an- chored targeting agent (TA) carrying the radionuclide (RN) emitting Cerenkov radiation and activating the PS and destroying the nano-carrier. The present method relates to internally stimulated in situ drug release based on the use of Cerenkov radiation generated within the body by intrave- nously administered medically used radionuclide ( s ) . Cerenkov radiation is used as a trigger for drug release rather than direct killing of cancer cells by radical formation as applied in prior art, which is more treatment-efficient. When using Cerenkov radiation as a trigger, the exact location of the nano-carrier in the cell (or on the cell) is found to be of lesser importance. While when relaying on radicals for cell killing their short half-life, especially of singlet oxygen, limits their diffusion distance, implying that for good cell killing efficiency they need to be very close to essential cell organelles (e.g. the cell nucleus). The present trigger approach is considered to rely on time sequential intravenous administration of a Cerenkov responsive drug nano-carrier and the trigger, i.e. a radionuclide coupled to a tumour-targeting agent. This concept is based on the use of a radionuclide generating Cerenkov radiation coupled to a targeting agent which when arriving at the tumour triggers release from nano-carriers already accumulated at the malignant site, Figure 1. Cerenkov radiation is generated as long as the radionuclide has not decayed. Based on the different bio-distribution of the radionuclide (renal clearance) and the drug carrier {hepatic clearance) , release can be facilitated only at the tumour site. This new trigger method does not depend on the characteristics of the local environment for release, allowing to choose the best time to trigger delivery. The design of a suitable nano-carrier for Cerenkov triggered release allows the use of diagnostic ra ^¬ dionuclides such as ⁶⁸ Ga at normally applied radioactivity levels, giving a patient radiation dose not higher than a X- ray CT scan, i.e. very low. It is noted that effectiveness of phototherapeutic application of Cerenkov radiation per se as initiated by radionuclides is questionable. It has been calculated that the produced energy and photon fluence rate for several radionuclides was in the order of nJ/cm ² . These results indicate than no therapeutic effects can be realised, possibly with the exception of cases where a high dose can be deposited at the tumour, and external radiotherapy due to higher photon fluence. It relates to a rather novel and original approach to tumor-targeted chemotherapy.

Therein aspects of PDT in combination with targeted tumor delivery to release chemotherapeut ic drugs only at the tumor are used, in particular enabling attack of deeply situated and unknown metastases with very limited side effects. Cerenkov radiation generated by radionuclides at the tumor is used as a source of UV-photons to activate chemotherapeut ic agents. The Cerenkov radiation is essentially harmless UV light such as created by energetic β-particles when transferring through a medium. The present Cerenkov radiation has a broad spectrum (X _ma x~360 nm) , which enables activation of UV active substances used in PDT. Certain β-particle emitting radionuclides can have a very high yield of Cerenkov photons and are even currently employed in Cerenkov preclinical imaging. In an example these radionuclides are delivered to a tumor site by conjugation to certain peptides. There, the generated Cerenkov photons can initiate release of a chemotherapeut ic agent. Radionuclides such as ⁶⁸ Ga, giving 34 photons per decay, may be used in diagnostics and show no adverse health effects (in fact they have a lower radiation dose than CT scans) . Two approaches are followed. In the first approach very stable nano-carriers such polymerases are used, where the UV active sensitizer is encapsulated or incorporated in the po- lymerosome membrane while in the lumen a drug is enclosed. When exposed to Cerenkov UV-photons, the sensitizer will produce highly reactive singlet oxygen species, which will destroy the membrane and therefore release the drug.

In the second approach so-called prodrugs are used, which are chemically inactivated drugs that can be activated by UV light. In their native form prodrugs are harmless and not effective. Reaction with Cerenkov radiation will locally activate the drug, killing the tumor cells. Potential prodrugs are O-nitrobenzyl-protected Doxorubicin and Auristatin E/F.

An important part of both approaches is that the drug release or activation is only triggered when exposed to the Cerenkov radiation coming from the radionuclide, which is located only at the tumor site. In both approaches, a combination of timed sequential administration of the radionu- elide and the drug, and a difference in bio distribution, ensures that the radionuclide and the drug only meet at the tumor. The radionuclide is in an example coupled to a peptide, which peptide may also be used in selective tumor visualization, which peptide is typically cleared from the body in a few hours and has no accumulation in liver or spleen

(clearance organs of nano-carriers) and short residence time in the kidneys. The nano-carrier or the prodrug is typically injected first, and is given sufficient time to achieve total elimination from the blood stream, after which the radi- onuclide bound to the peptide will be administered.

EXAMPLES/EXPERIMENTS

The invention although described in detailed explanatory context may be best understood in conjunction with the accompanying examples.

Production of singlet oxygen

A singlet oxygen ¹ 02 is produced by ¹⁶⁶ Ho and ⁶⁸ Ga .

In contrast to normal ground state oxygen, ¹ 02 possesses a high reactivity and may readily react with unsaturated organic molecules. The reactivity of ¹ 02 is found to strongly depend on its lifetime, which is mainly controlled by solvent properties; for instance in water a kinetic lifetime ΤΔ of ~ 3.3 is has been reported.

In general ¹ 0? can be produced via photochemical pathways by means of a photosensitizer (PS) molecule or via chemical sources, for example reaction of sodium hypochlorite with hy ^¬ drogen peroxide. Suitable PSs are for instance tetraphenylpor- phyrin, Methylene blue and Rose Bengal. In the experiments further Phenalenone and Chlorin e6 were used. Phenalenone has an outstanding ^Ύ θ2 quantum yield of 0.99 (¾0, 337-439 nm) and an extinction coefficient of 11250 M ^_1 cm ^"1 (MeOH, 360 nm) .

Chlorin e6 has a smaller ^x 02 quantum yield of 0.65 (EtOH, 347 nm) but a large extinction coefficient of 200000 M ^_1 cm ^-1 (EtOH, 401 nm) . With regard to the Cherenkov spectrum, which has the highest intensity between 300 and 400 nm, Phenalenone covers a wider range of the radiation than Chlorin e6. Furthermore, the ¹ 02 quantum yield of Phenalenone is wavelength-independent over a broad region (337-439 nm) .

Materials and methods

Chlorin e6 was purchased from Frontier Scientific. SOSG (Singlet Oxygen Sensor Green) was purchased from Thermo Fisher. Phenalenone, Acetonitrile , HEPES (4- (2-hydroxyethyl) pipera- zine-l-ethanesulfonic acid) and HoCl3-6H20 (Holmium(III) chloride hexahydrate) were purchased from Sigma Aldrich. EDTA (Ethylenediaminetetraacetic acid) was purchased from Merck.

Ultrapure water was prepared with the in-house Milli-Q system from Merck Millipore. Measurements were conducted in UV transparent cuvettes by means of a Cary Eclipse Fluorescence Spectrophotometer from Agilent Technologies with the following settings: Velocity Fast, Medium detector voltage, KE _X = 509 nm for SOSG*. The LED (400 nm) was purchased from Ledshighpower (Landgraaf, The Netherlands). In general, all operations were carried out under dark conditions.

Preparation of PSs

Glass vials were wrapped in aluminium foil and sealed with caps. Phenalenone was first dissolved in methanol and vortexed for 1 minute. Afterwards, it was further diluted in HEPES (100 mM) to reach a final concentration of 1 mM with 10% (v/v) methanol. Chlorin e6 was dissolved in HEPES (100 mM) to reach a final concentration of 100 μ . Afterwards, it was put in the ultrasonic bath for 30 minutes. In the experiments, the PSs were further diluted with HEPES (100 mM) to reach the desired concentration

Preparation of Singlet Oxygen Sensor Green <SOSG)

SOSG was dissolved in methanol to reach a final concentration of 1.2 mM. In the experiments, SOSG was further diluted with HEPES (100 mM) to reach a final concentration of 6 μΜ.

Chlorin e6 exposed to LED light

2 ml of Chlorin e6 (10, 15, 20, 25, 30, 40 or 50 μΜ) together with SOSG were placed in a cuvette, subsequently sealed with a cap and exposed to the light source (2.8 V, 0.75 A) mounted inside the fluorimeter.

PSs exposed to Ho-166

2 ml of Chlorin e6 (20 μΜ) and Phenalenone (100 μΜ) , respectively, together with SOSG were placed in a cuvette and positioned in the closed fluorimeter. HoCl ₃ -6H20 (3 mg) was activated in a nuclear reactor (Reactor Insitute Delft, The Netherlands, thermal neutron flux 3.1 X 10 ¹⁶ n s" ¹ rrr ² , 10 hours). The activated Holmium salt (200 Bq) dissolved in 200 μΐ EDTA solution (65.9 mM in 100 mM HEPES) was added into the cuvette and sealed with a cap. Controls contained the same amount of non-activated Holmium salt.

Phenalenone exposed to Ga-68

2 ml of Phenalenone (100 μΜ) together with SOSG were placed in a cuvette and positioned in the closed fluorimeter. Ga-68 was eluted from a Ge-68/Ga-68 generator (Eckert & Ziegler) with 0.1 M HC1 in fractions of 0.5 ml. The fraction containing the highest activity (65 MBq) was added to a solution consisting of 500 μΐ HEPES (100 mM) , 50 μΐ NaOH (1 M) and 10 μΐ EDTA (65.9 mM in 100 mM HEPES) .

The final mixture was added into the cuvette and sealed with a cap .

Results and Discussion

Chlorin e6 exposed to LED light

Chlorin e6 was exposed to LED light in the presence of SOSG. The production of ^λ θ2 was tracked by measuring generated SOSG* via fluorescence spectrometry. It was found that a higher pH led to a decreased production of ^λ θ2. A control only containing SOSG without any PS did not cause any production of ¹ θ ₂ · It is noted that the reduced production of the oxygen spe ^¬ cies is attributed to the SOSG molecule itself and not to the PS. The fluorescence of the SOSG-probe is found very sensitive to pH changes. The highest production of ¾2 was found within a range of 15 - 20 μΜ PS. Further increase of the concentration led to aggregation of Chlorin e6, accompanied by a drop in production of the oxygen species. Chlorin e6 molecule starts to aggregate within the range of 15.1 - 16.8 μΜ at pH 7.

Therefore, in all the following experiments a PS concentration of 20 μΜ at a pH of 7. was used.

Singlet oxygen produced by means of radioisotopes

Photosensitizers were exposed to beta emitting radioisotopes to investigate whether the Cherenkov radiation is sufficient enough to produce ² θ2. The production of the oxygen species was detected by means of SOSG. Generated SOSG* was measured using fluorescence spectrometry. During the research, it was noted that SOSG* completely disappeared in the presence of Ho ³⁺ and could not be detected with the fluorimeter anymore. Only when the holmium salt was chelated with EDTA, SOSG* could still be measured .

Phenalenone and Chlorin e6 in the presence of SOSG were exposed to Ho-166. It was found that ^χ θ2 was produced, indicated by an increase of the intensity maximum of SOSG*. Controls containing no radioisotope but only the respective PS together with SOSG show that the absence of Cherenkov radiation did not lead to any oxygen species. Comparing the production rate of ^x 02 between Phenalenone (orange) and Chlorin e6 (blue) at 180 minutes, more than twice the amount of singlet oxygen was pro- duced by Phenalenone. This can be explained by the high ¹ 0 quantum yield of Phenalenone (Φ « 1.0} and its absorption spectrum which covers a wider range of the Cherenkov radiation than Chlorin e6. In addition, Phenalenone was applied in a higher concentration than Chlorin e6 since it is not con- strained to a certain concentration maximum. Higher concentrations (>100 μΜ, e.g. up to 500 μΜ) Phenalenone can be employed to increase production of ¹ 02.

Besides Ho-166, Phenalenone was exposed to the radioisotope Ga-68. In contrast to the beta minus emitting Ho-166, Ga-68 decays via beta plus emission. With regard to the photodynamic properties,, Ga-68 emits 33.9 photons per decay, while Ho-166 only emits 27.5 photons per decay. The intensity of Ho-166 re ^¬ mains longer since its half-life time is almost 27 hours, while the half-life time of Ga-68 is only 67 minutes. It was found that the exposure of the PS to Ga-68 led to a production of the oxygen species, indicated by an increase of the intensity maximum of SOSG* . Comparing the ¾ production caused by Ho-166 and Ga-68 at 150 minutes, more than twice the amount of ¹ 0 ₂ was produced by Ho-166. This can be explained by the difference in activities. Ga-68 was eluted from a Ge-68/Ga-68 generator with a final activity of 65 MBq, while the Holmium salt activated in the reactor had an activity of 200 MBq. The higher the activity, the more beta particles are emitted over time and the stronger the intensity of the Cherenkov radiation. Therefore, 200 MBq of Ho-166 led to a higher production rate of ^ ₂ than 65 MBq Ga-68.

Loading and release of chlorine e6.

Loading of chlorin e6 has been carried out in micelles com- posed of PEO-PB (polyethylene oxide-polybutadiene 400-1800) . Table 1 shows the achieved loading efficiencies for different formulations .

Table 1: Loading efficiencies of 20 μΜ (final concentration) Ce6 in PB-PEO. Ce6 was dissolved in chloroform or acetone and loaded in micelles simultaneously with the micelle self- assembly process using a combination of an oil-in-water method and a co-so-lvent evaporation method. The stock solution may be used to obtained diluted solutions.

Polymer Stock

M concentration Loading ef

Polymer (g/mol) (mg/ml) Cosolvent ficiency

PB-PEO ¹ 1800-4000 200 chloroform 93 + 5%

PB-PEO ² 1800-4000 200 acetone 96 ± 0.3%

PB-PEO ² 5200-4500 50 chloroform 81 ± 12%

PB-PEO ² 5200-4500 200 chloroform 81 ± 10%

Figure 2 shows the release of Chlorin e6 (in terms of retention) as a function of radiation dose delivered by a Co-60 source in micelles composed of PB-PEO (1800-400). The error bars correspond to uncertainties determined by three separate experiments. The line is an exponential fit of the data. Co-6 decays by beta minus emission and emits also two highly ener ^¬ getic gamma rays which through the photoelectric and Compton effect free electrons resulting in the creating of Cerenkov radiation. Chlorin e6 is a photosensitizer that when activated by light produces singlet oxygen. Chlorin e6 is itself fluorescent and can be used to follow its release from the micelles when irradiated by gamma's, which fluorescent is there ^¬ fore an indirect way of monitoring the release of a payload. Chlorin e6 is very hydrophobic and when not exposed to radia- tion it is not released from the micelles at all. X-ray irradiation at energy below the Cerenkov threshold does not lead to release either, indicating that singlet oxygen production through Cerenkov light must play a role in the release.

So fig. 2 shows the retention of Chlorin e6 as function of the radiation dose. It has been found that Chlorin e6 is loaded with different efficiency in different polymeric carriers, see table 1. For the retention experiments the polymer PB-PEO {1800-400) was used. The intrinsic release of Chlorin e6 from the micelles (i.e. the release in the absence of radiation) has been determined to be close to zero and may therefore be considered to be "absent". As a fact inventors did not measure any release. This can be explained by the lipophilic character of chlorin e6, which even if released will likely go back, to the hydrophobic part of the micelles. This is also in line with the high loading efficiencies found. Figure 2 demonstrates that apparently there is an exponential release of chlorin e6 as function of dose. In this experiment chlorin e6 was used as the generator of singlet oxygen (i.e. photosensitizer) responding to Cerenkov light, as well as a 'dye' to evaluate the release of the system. It has to be understood that upon production of the singlet oxygen a chemotherapeutic drug present in the vehicle (micelles) being usually less lipophilic are released in larger amounts compared to chlorin e6. To evaluate the killing efficiency of the present system, the vehicles, such as the micelles, are loaded (10~ ⁴ , 10 ^-3 , 10 ^-2 , and 10 ^'1 gr/1) with the photosensitizer (10" ⁶ , 10 ^~4 , 10 ^"2 , and 10 ^"1 gr/gr micelle) and the chemotherapeutic drug simultaneously (lO ^"5 , 10 ^"3 , 10 ^"2 , and lO ^"1 gr/gr micelle) . Afterwards the loaded micelles are added to various cell cultures, preferably tumour spheroids, such as MDA- B-231 and LNCaP, and subsequently exposed to radiation, such as gamma radiation at different doses (0.1-80 GY) , such as with ⁶⁸ Ga, ⁹⁰ Y, ⁶⁰ Co, ¹⁷⁷ Lu, and ¹⁶⁶ Ho, coupled to an antigen, such as a prostate specific membrane antigen, such as Glu-NH-CO-NH-Lys- (Ahx) - ⁶⁸ Ga- (HBED- CC) , and PSMA-617. Exposure times range for example between 3- 15 h, at environmental temperatures, such as room temperature. An impact on the cultures is measured, such as by measuring fluorescence, after 1 day, 3 days, 7 days and 3 weeks. There- after cell culturing is stopped. Typically multiwall plates were used and experiments were carried out at least in duplicate, and typically at least in triplicate. When chlorin e6 was present no staining was needed, as the chlorin e6 was detectable using fluorescence by itself. Experiments on both 2D and 3D tumour spheroid models are carried out. Confocal microscopy is applied to evaluate the distribution of carriers throughout the spheroids. For this purpose, thin slices of 20 μπι are made and analysed. In control experiments empty micelles are used, micelles containing chlorin e6, and micelles loaded only with a chemotherapeutic drug (such as docetaxel and resveratrol) (in the same amounts as above for the loaded variants) . In addition the growth of the tumour spheroids are evaluated for different radionuclide concentrations (10 ^~10 , 10 ^{~ 8} , 10 ^"6 , and 10 ^"1 gr/1) and targeting agent (10 ^~14 , 10' ¹² , 10" ¹⁰ , 10 ^"s , mole/1) . The micelles containing chlorin e6 and the chemotherapeutic drug lead at least to a reduced growth and typically even to shrinkage of the tumours at a much lower dose than the control experiments, typically 5-50% lower dose or more.

Experiments are further conducted on smaller animals.