Composition for the treatment or prevention of unwanted immune responses

Abstract

The invention relates to a composition for use in the treatment or prevention of one or more unwanted immune responses in a subject, wherein the one or more unwanted immune responses are caused by the use of one or more immunogenic drugs and wherein the composition comprises negatively charged non-polymeric nanoparticles, wherein the negatively charged non-polymeric nanoparticles are selected from the group consisting of at least one of silicon particles, solid lipid particles comprising at least one immune modulating drug and liposomes comprising at least one immune modulating drug.

Description

[0001] 91.168061 / 01

[0002] Composition

[0003] Field of Invention

[0004] The present invention relates to a composition for use in the treatment or prevention of one or more unwanted immune responses in a subject, wherein the one or more unwanted immune responses are caused by the use of one or more immunogenic drugs and wherein the composition comprises negatively charged non-polymeric nanoparticles wherein the negatively charged non-polymeric nanoparticles are selected from at least one of silicon particles, solid lipid particles comprising at least one immune modulating drug and liposomes comprising at least one immune modulating drug.

[0005] Background

[0006] Pharmaceutical compositions comprising macromolecular immunogenic drug substances or other components often possess side effects (for example due to their immunogenicity) and reduced therapeutic efficacy due to these drug substances having unwanted immunogenic properties causing unwanted immune responses. During the last 20 years, many macromolecular drug substances have been developed and approved by regulatory bodies for the treatment of various diseases. These drug substances are typically protein-based compounds and compounds for gene therapy. Macromolecular drug substances are typically compounds with a molecular weight above 5 kDa; for example, monoclonal antibodies and nucleic acid-based drug substances with molecular weight above 100 kDa.

[0007] Clinical data have shown that the therapeutic potential of such drugs is much greater than the therapeutic potential of traditional low molecular drug substances. However, the clinical use of macromolecular drugs is often associated with several additional challenges when compared to the clinical use of small molecules. One potential problem related to clinical use of macromolecular drugs is the occurrence of unwanted immune reactions that may limit the clinical value of the drug. In general, the risk factors for such unwanted immune reactions are both patient related and drug related. Macromolecular drug substances that are chemically very different from endogenous compounds (i.e. compounds produced inside an organism or cell) may have a greater potential for unwanted immunological reactions than drugs that are 91.168061 / 01 endogenous or chemically similar to natural compounds. In addition, multiple administrations of the same immunogenic drug over time may potentiate unwanted immune responses. Such immunological reactions may affect the pharmacokinetic properties of the immunogenic drug, resulting in side-effects and thereby raising patient safety issues and may also reduce the treatment efficacy of the macromolecular drug. Several macromolecular drugs are currently under development and unwanted immunological reactions may be a critical issue for the ultimate clinical use of these agents.

[0008] The unwanted immunological reactions are often described as anti-drug antibody responses (AD As) because the formation of antibodies is a central element in the immune reaction. AD As are a major problem for many drug substances. Examples of drug substances which can trigger AD As, within the field of oncology, include the monoclonal antibodies nivoumab, trastuzumab, trastuzumab emtasine, elotuzumab, dinutuximab, catumaxomab, tositumomab and brentuximab vedotin (See Van Brummelen et al., The Oncologist, Vol. 21, Issue 10, October 2016, 1260-1268).

[0009] The immunological problem is also present for other macromolecular drugs (for example, fusion proteins) for other indications as well as for other drug substances larger than proteins and nucleic acids (for example, antisense nucleotides, RNA therapeutics and viral vectors). Particulate drug formulations for intravenous use may also result in unwanted immunological reactions.

[0010] Several immune modulating drugs (also known as “immunomodulatory drugs”) have been suggested for the improvement of unwanted immune reactions associated with administration of macromolecular drugs. However, use of these immune modulating drugs per se results in general immune suppression, and these drugs cannot be used for a long period of time because of the accompanying side effects.

[0011] WO2012149454 (Selecta Bioscience) describes synthetic nanocarrier compositions, and related methods, comprising CD Id-restricted antigens and immunosuppressants that provide tolerogenic immune responses. This patent document focusses on the use of biodegradable polymer particles comprising rapamycin to prevent or reduce immune responses / AD As related to clinical use of macromolecular drugs. However, prophetic Example 2 in WO2012149454 describes gold-decorated silica particles with coupled ibuprofen and prophetic Example 3 in WO2012149454 describes liposome-containing cyclosporine A as a 91.168061 / 01 prophetic product. The liposomal product comprises DPPC and cholesterol resulting in a neutral or positively charged liposome composition.

[0012] Summary of the invention

[0013] In a first aspect, the invention provides a composition for use in the treatment or prevention of one or more unwanted immune responses in a subject, wherein the one or more unwanted immune responses are caused by the use of one or more immunogenic drugs and wherein the composition comprises negatively charged non-polymeric nanoparticles, wherein the negatively charged non-polymeric nanoparticles are selected from the group consisting of at least one of silicon particles, solid lipid particles comprising at least one immune modulating drug and liposomes comprising at least one immune modulating drug.

[0014] In a second aspect, the invention provides the use of a composition for the manufacture of a medicament for the treatment or prevention of one or more unwanted immune responses in a subject, wherein the one or more unwanted immune responses are caused by the use of one or more immunogenic drugs and wherein the composition comprises negatively charged non- polymeric nanoparticles, wherein the negatively charged non-polymeric nanoparticles are selected from the group consisting of at least one of silicon particles, solid lipid particles comprising at least one immune modulating drug and liposomes comprising at least one immune modulating drug.

[0015] In a third aspect, the invention provides the use of a composition in the treatment or prevention of one or more unwanted immune responses in a subject, wherein the one or more unwanted immune responses are caused by the use of one or more immunogenic drugs and wherein the composition comprises negatively charged non-polymeric nanoparticles, wherein the negatively charged non-polymeric nanoparticles are selected from the group consisting of at least one of silicon particles, solid lipid particles comprising at least one immune modulating drug and liposomes comprising at least one immune modulating drug.

[0016] In a fourth aspect, the invention provides a method of treating or preventing one or more unwanted immune responses in a subject, wherein the one or more unwanted immune responses are caused by the use of one or more immunogenic drugs, wherein the method comprises the administration of a composition comprising negatively charged non-polymeric 91.168061 / 01 nanoparticles, wherein the negatively charged non-polymeric nanoparticles are selected from the group consisting of at least one of silicon particles, solid lipid particles comprising at least one immune modulating drug and liposomes comprising at least one immune modulating drug.

[0017] Definitions

[0018] The term “particle” means any form of water-insoluble material; typically silicon-based particle, lipid-based particle and liposomes.

[0019] The term “negatively charges particles” means particles that have an overall total negative charge. The negative particle charge is a based on the presence of acidic functional groups and can be typically determined by zeta potential measurements at pH 7.4 and temperature 37 degrees Celsius. The method for determination of zeta potential is well-known in the art. See for example Johnsen, HM et al. in International Journal of Pharmaceutics, 629 (2022), 122371

[0020] The term “nanoparticles” means particles that have an overall size less than 400 nm; typically for example 30-300 nanometer, preferably 100 to 290 nm using the technique of Dynamic Light Scattering (DLS), for example using instruments like Zetasizer.

[0021] The term “non-polymeric” means that the composition of the particles is not made of polymeric material, for example polylactic acid. The particles may however comprise one or more polymers as surface components. For example, polyethylene glycol (PEG) may be present as a surface component non-covalently or covalently bond to the non-polymer particle surface.

[0022] The term “macromolecular drugs” means drug substances with a molecular weight at or above 5000 Da, typically 20 000 to 150 000 Da.

[0023] The term “protein drugs” means any protein or derivative thereof that can be used for the treatment of disease.

[0024] The term “protein conjugates” means a protein comprising additional other components than amino acids. Typical protein conjugates include a protein where a biologically active substance or polyethylene glycol is covalently coupled to the protein by a covalent bond. The 91.168061 / 01 protein conjugate might comprise a spacer between the protein and the biologically active substance or polyethylene glycol.

[0025] The term “subject” means humans or animals, including warm blooded mammals such as humans and primates; avians; domestic household or farm animals such as cats, dogs, sheep, goats, cattle, horses and pigs; laboratory animals such as mice, rats and guinea pigs; fish; reptiles; zoo and wild animals; and the like. Preferably the subject is a human subject.

[0026] The term “solid lipid” means a nanoparticle having a solid wax or lipid core surrounded by a layer of surfactants.

[0027] The term “liposome” means having one or more rings of a lipid bilayer surrounding an interior space, generally wherein the interior space is aqueous.

[0028] The term “pharmaceutical formulation” includes “drug product” and refers to a composition comprising at least one drug substance and at least one excipient.

[0029] The term “drug” or “drug substance” as used herein refers to any biologically and / or pharmacologically active compound including prodrugs thereof. Any stereoisomer, or pharmaceutically acceptable salt or solvate thereof are included in the present term. The term drug substance includes any drug substance with regulatory approval, drug substances in current development and drug substances that have been on the market.

[0030] The term “pharmaceutically acceptable” refers to chemical compounds and mixtures thereof that are acceptable to be used in drug products. All excipients used in regulatory approved drug products are pharmaceutically acceptable.

[0031] The term “excipient” refers to chemical compounds for use in drug products where said excipients per se are not biologically active in the amount present when the drug product is used according to the intension or regulatory approval.

[0032] The term “silicon zero comprising particles” refers to particles where at least 50% of the present silicon is with oxidation level zero and not four as in silica.

[0033] The term “immunogenic drug” refers to drugs and drug substance which are capable of provoking an immune response in the body of a subject wherein the term “subject” is as hereinbefore defined. Wherein the one or more unwanted immune responses is specifically 91.168061 / 01 an ADA response, the immunogenic drug may also be herein referred to as an “ADA forming drug”.

[0034] Detailed description of the invention

[0035] As would be understood by the skilled person, any of the aspects disclosed herein may be combined with one another to arrive at further aspects of the invention. Furthermore, it will be appreciated that any aspect disclosed herein in the context of a “composition for use” is also equally applicable to the “use” and “method” embodiments described above in the summary of invention.

[0036] The present invention relates to a composition for use in the treatment or prevention of one or more unwanted immune responses in a subject, wherein the one or more unwanted immune responses are caused by the use of one or more immunogenic drugs and wherein the composition comprises negatively charged non-polymeric nanoparticles.

[0037] In the context of the present invention, by “treatment” and “prevention” we typically mean the reduction of the one or more unwanted immune responses. By “unwanted immune response” we mean, for example, complement mediated inflammation, cytokine storm, cell and tissue toxicity, thrombotic microangiopathy, autoimmune disease, transplant rejection, and allergic reactions. In particular, we mean anti-drug-antibody (ADA) responses and other undesired T- and / or B-cell mediated immune responses. Thus, the compositions for use in the present invention are preferably for use in the treatment or prevention of one or more ADA responses in a subject.

[0038] The formation of ADA is detected for very many macromolecular drugs including antibodies. See for example Van Brummelen, E.M.J. et al. in The Oncologist 2016,21,1260-1268 and references herein.

[0039] A preferred aspect of the present invention is to reduce or eliminate AD As formed by administration of drugs comprising biological drug substances or particulate (water-insoluble) drug formulations.

[0040] A preferred aspect of the present invention is to reduce or eliminate AD As formed by administration of drugs comprising monoclonal antibodies or conjugates thereof. 91.168061 / 01

[0041] Nanoparticles

[0042] In certain aspects, the non-polymer nanoparticles are silicon particles.

[0043] In other aspects, the non-polymer nanoparticles are solid lipid particles comprising at least one immune modulating drug.

[0044] In further aspects, the non-polymer nanoparticles are liposomes comprising at least one immune modulating drug.

[0045] In certain aspects, the negatively charged non-polymeric nanoparticles have a zeta potential below minus 10 mV in pure water at 37 degrees Celsius.

[0046] In certain aspects, the negatively charged non-polymeric nanoparticles have an average particle size of less than 400 nm, using the technique of Dynamic Light Scattering (DLS), for example using instruments like Zetasizer.

[0047] In certain aspects, the one or more immunogenic drugs causing the unwanted immune response is a protein drug substance or nucleic acid-based drug substance, including viral vehicles such as an adenovirus associated virus carrying a drug substance (e.g. a therapeutic gene).

[0048] In certain aspects, the one or more immunogenic drugs causing the unwanted immune response is in the form of a water-insoluble particulate.

[0049] In certain aspects, the one or more immunogenic drugs comprise or consist of macromolecular drug substances. Preferably or alternatively, the one or more immunogenic drugs are selected from drugs for treatment of cancer, autoimmune diseases, respiratory diseases, gastrointestinal diseases, cystic fibrosis, cardiovascular disease, central nervous system diseases, diabetes, hemophilia, AIDS and eye diseases.

[0050] In certain aspects, the one or more immunogenic drugs comprise or consist of particulate drug formulations; including for example liposome or lipid formulations.

[0051] In certain aspects, the negatively charged non-polymeric silicon nanoparticles are free from any drug or drug substance. 91.168061 / 01

[0052] In other aspects, the negatively charged silicon non-polymeric nanoparticles are free from any drugs other than the one or more immunogenic drugs causing the unwanted immune response and optionally one or more immune modulating drug substances triggering immune tolerance.

[0053] In other aspects, the negatively charged non-polymeric nanoparticles comprise the one or more immunogenic drugs, or fragments thereof, causing the unwanted immune response.

[0054] In other aspects, the negatively charged non-polymeric silicon nanoparticles comprise one or more immune modulating drug substances.

[0055] In one preferred aspect of the present invention, the negatively charged non-polymeric nanoparticles are silicon nanoparticles and the negatively charged non-polymeric nanoparticles comprise rapamycin or everolimus, optionally wherein the rapamycin or everolimus is in the form of a complex with 2-hydroxypropyl-beta-cyclodextrin or sulfobutyl ether beta-cyclodextrin including pharmaceutically acceptable salts thereof .

[0056] In another preferred aspect of the present invention, the negatively charged non-polymeric nanoparticles are silicon nanoparticles and the negatively charged non-polymeric nanoparticles comprise rapamycin or everolimus, optionally wherein the rapamycin or everolimus is in the form of a complex with 2-hydroxypropyl-beta-cyclodextrin or sulfobutyl ether beta-cyclodextrin including pharmaceutically acceptable salts thereof.

[0057] In another preferred aspect of the present invention, the negatively charged non-polymeric nanoparticles are liposomes comprising at least one immune modulating drug and the negatively charged non-polymeric nanoparticles comprise rapamycin or everolimus, optionally wherein the rapamycin or everolimus is in the form of a complex with 2- hydroxypropyl-beta-cyclodextrin or sulfobutyl ether beta-cyclodextrin including pharmaceutically acceptable salts thereof.

[0058] In another preferred aspect of the present invention, the negatively charged non-polymeric nanoparticles are lipid nanoparticles comprising at least one immune modulating drug and the negatively charged non-polymeric nanoparticles comprise rapamycin or everolimus, optionally wherein the rapamycin or everolimus is in the form of a complex with 2- hydroxypropyl-beta-cyclodextrin or sulfobutyl ether beta-cyclodextrin including pharmaceutically acceptable salts thereof. 91.168061 / 01

[0059] In another preferred aspect of the present invention, the negatively charged non-polymeric nanoparticles are silicon nanoparticles and the negatively charged non-polymeric nanoparticles comprise both rapamycin or everolimus, optionally wherein the rapamycin or everolimus is in the form of a complex with 2-hydroxypropyl-beta-cyclodextrin or sulfobutyl ether beta-cyclodextrin including pharmaceutically acceptable salts thereof and the ADA forming drug; preferably a monoclonal antibody, a protein conjugate, including a monoclonal antibody or pharmacological active peptides and peptidomimetics.

[0060] In another preferred aspect of the present invention, the negatively charged non-polymeric nanoparticles are liposomes comprising at least one immune modulating drug and the negatively charged non-polymeric nanoparticles comprise both rapamycin or everolimus, optionally wherein the rapamycin or everolimus is in the form of a complex with 2- hydroxypropyl-beta-cyclodextrin or sulfobutyl ether beta-cyclodextrin including pharmaceutically acceptable salts thereof and the ADA forming drug; preferably a monoclonal antibody a protein conjugate, including a monoclonal antibody or pharmacological active peptides and peptidomimetics.

[0061] In another preferred aspect of the present invention, the negatively charged non-polymeric nanoparticles are lipid nanoparticles comprising at least one immune modulating drug and the negatively charged non-polymeric nanoparticles comprise both rapamycin or everolimus, optionally wherein the rapamycin or everolimus is in the form of a complex with 2- hydroxypropyl-beta-cyclodextrin or sulfobutyl ether beta-cyclodextrin including pharmaceutically acceptable salts thereof and the ADA forming drug; preferably a monoclonal antibody a protein conjugate, including a monoclonal antibody or pharmacological active peptides and peptidomimetics

[0062] The negatively charged non-polymeric nanoparticles typically have a zeta potential below -10 mV in pure water at 37 degrees centigrade. Even more preferably, the zeta potential is below -20 mV in pure water at 37 degrees centigrade. Even more preferably, the zeta potential is below -30 mV in pure water at 37 degrees centigrade.

[0063] In one aspect of the present invention, the negatively charged non-polymeric nanoparticles have an average size less than 400 nm, using the technique of Dynamic Light Scattering (DLS), for example using instruments like Zetasizer. Preferably, the negatively charged non- 91.168061 / 01 polymeric nanoparticles have an average size less than 350 nm, more preferably less than 320 nm, for example 50 to 300 nm, such as 100 to 290 nm.

[0064] Silicon

[0065] In one aspect of the present invention, the negatively charged non-polymeric nanoparticles are silicon nanoparticles.

[0066] The silicon in the silicon nanoparticles of the present invention is present in at least 50 wt% as elemental silicon (silicon with oxidation number 0), relative to the total weight of silicon. More preferred form of silicon in the present silicon particles is at least 70 wt% as elemental silicon, even more preferred at least 80 wt% as elemental silicon, relative to the total weight of silicon. Another preferred aspect related to the form of silicon in the present particles is that the amount of elemental silicon is more than 80%, more preferably more than 90% most preferably more than 95%, relative to the total weight of silicon.

[0067] In another preferred aspect of the present invention, the silicon nanoparticles do not comprise more than 20 wt% silicon with oxidation level above zero relative to the total weight of the silicon nanoparticles.

[0068] In another preferred aspect of the present invention, the silicon nanoparticles are produced by chemical vapor deposition (CVD).

[0069] A CVD process is a process wherein a gas is converted to a solid material, typically a film, under various conditions. Step a) of the process of the invention preferably involves preparing silicon particles via CVD from a silicon containing reaction gas, such as silane or trichlorosilane, preferably silane gas.

[0070] In particular, the silicon particles produced by CVD are collected by a centrifugal unit in the reactor. Preferably, the CVD process is carried out in a reactor wherein the reactor comprise a reactor body and a rotation device operatively arranged to the reactor, wherein the rotation device is configured to rotate the reactor around an axis during production of said silicon comprising particles; hereafter referred as cCVD-SP (centrifuge Chemical Vapor Deposition Silicon Particles). 91.168061 / 01

[0071] In a further preferred aspect of the present invention, the CVD process is carried out in a reactor wherein the reactor comprise a reactor body and a rotation device operatively arranged to the reactor, wherein the rotation device is configured to rotate the reactor around an axis during production of said silicon comprising particles; hereafter referred as cCVD-SP, followed by an etching process to prepare the porosity of the particles. Such particles are here referred to as PcCVD-SP (Porous centrifuge Chemical Vapor Deposition Silicon Particles).

[0072] The etching process for production of PcCVD-SP from cCVD-SP could be similar to other well-known etching processes of silicon particles described in the prior art; for example, a hydrofluoric acid based method.

[0073] Preferably, the present PcCVD-SP particles are produced by an aggregation process taken place in a reactor, forming stable aggregates and thereby porosity without any etching. All silicon particles in the examples are produced by this non-etching process.

[0074] The particle surface may be modified to exhibit desired characteristics; including chemical or thermal oxidation or coating.

[0075] The elemental silicon in the particles of the invention may be in amorphous or crystalline form. The elemental silicon in particles produced by the CVD process is mainly in the form of amorphous elemental silicon at ambient temperature, however, particles comprising crystalline silicon can directly be prepared by CVD at high temperature (e.g, 600 °C and above) and longer reaction times. The particles comprising crystalline silicon prepared from a CVD method typically are in the form of polycrystalline material (crystal size around 1.5 nm) while crystalline milled particles typically consist of one crystal of silicon.

[0076] The crystalline versus amorphous form of silicon can routinely be determined by X-ray diffraction analysis (XRD analysis). The amorphous form of silicon can be transformed to crystalline form of silicon by heating to relative high temperatures (e.g. above 500 °C).

[0077] Silicon particles produced by the CVD method typically comprise some material comprising one or more silicon-hydrogen bond. This hydrogen might be available for formation of some hydrogen gas in a reaction with water.

[0078] In certain embodiments, the elemental silicon is present in a crystalline form, in some embodiments typically more than 50 wt% in the crystalline form and in some embodiments 91.168061 / 01 more than 70 wt% in a crystalline form and finally in some embodiments more than 90 wt% in a crystalline form, relative to the total weight of elemental silicon.

[0079] In other embodiments, the silicon particles comprise elemental silicon in amorphous form, in some embodiments more than 50 wt%, in some embodiments more than 70 wt%, in some embodiments more than 90 wt% and finally in some embodiments more than 95 wt% in amorphous form, relative to the total weight of elemental silicon.

[0080] Typical median diameters for the silicon particles of the invention may be less than 400 nm, such as 30 to 300 nm, using the technique of Dynamic Light Scattering (DLS), for example using instruments like Zetasizer. The given particle sizes are related to the final silicon particles loaded with one or more drug substances and optionally excipients and coating.

[0081] The poly dispersity index can also vary from almost monodisperse particles to particles with very broad particle size distribution.

[0082] The preferred particle size of the silicon particles of the invention will generally vary depending upon indication and route of administration. The particles can be administered by any known route of administration; including both enteral and parenteral administration.

[0083] The preferred routes of administration are oral administration, intravenous administration and subcutaneous administration. For silicon nanoparticles without any drug substances the most preferred routes of administration are orally in the form of capsules or tablets or by subcutaneous administration in the form of an injection.

[0084] For particles comprising immune suppressive drug substances like for example rapamycin, the most preferred route of administration is by subcutaneous administration in the form of an injection.

[0085] The silicon particles of the invention can be non-porous (cCVD-SP) or porous (PcCVD-SP). The most preferred particles according to the present invention are porous particles.

[0086] The function of the negatively charged non-polymeric nanoparticles administered parenterally is to target the immune modulating drug to the dentritic cells and thereby to the immune system to reduce the unwanted ADA effect. 91.168061 / 01

[0087] Without wishing to be bound by any theory, the mechanism behind the observed immune modulating effect of silicon particles without any immune modulating drugs may be that this is an effect of hydrogen gas generated when silicon (oxidation level zero) reacts with water.

[0088] The porosity of the PcCVD-SP can vary over a large range depending upon choice of drug substance, indication and administration route. The porosity is a measure on the volume of the pores. A PcCVD-SP with porosity of 50 % has a porosity volume that is 50% of the total PcCVD-SP volume. The porosity of PcCVD-SP may typically be from 20% to 90%. In certain embodiments, the porosity is more than 40%, typically more than 50%, more than 60%, more than 70%, more than 80% such as 90%.

[0089] The pore size of PcCVD-SP can vary from microporous particles through mesoporous particles to macroporous particles depending on nature of the drug substance, dose of the drug substance, indication, form of the drug product and route of administration. Typical average pore size of PcCVD-SP for loading of drug substances is from 1 nm to 200 nm. In one embodiment of the present invention, the average pore size is 1-10 nm, in another embodiment the typical pore size is 5-20 nm, in still another embodiment, the typical pore size is 10-50 nm and finally, in still another embodiment, the typical pore size is 2-50 nm.

[0090] The smaller pore size (a few nanometers) fits low molecular drug substances which have a size of about 1 nanometer or above, while larger pore size (20 nanometers or more) fits larger drug molecules like monoclonal antibodies which typically have sizes above 10 nanometers.

[0091] In one embodiment, the particles are microporous. In this embodiment, preferably at least 2 vol% of the pores are micropores, more preferably at least 5 vol%, even more preferably at least 10 vol%, especially at least 20 vol%, such as at least 50 vol%, relative to the total pore volume.

[0092] In another embodiment, the particles are mesoporous. In this embodiment, preferably at least 2 vol% of the pores are mesopores, more preferably at least 5 vol%, even more preferably at least 10 vol%, especially at least 20 vol%, such as at least 50 vol%, relative to the total pore volume.

[0093] In a further embodiment, the particles are macroporous. In this embodiment, preferably at least 2 vol% of the pores are macropores, more preferably at least 5 vol%, even more 91.168061 / 01 preferably at least 10 vol%, especially at least 20 vol%, such as at least 50 vol%, relative to the total pore volume.

[0094] In a preferred aspect of the present invention, the negatively charged non-polymeric nanoparticles are silicon nanoparticles wherein the silicon nanoparticles are free from any drugs. This aspect of the invention relates to plain silicon particles without any drug substances.

[0095] In another preferred aspect of the present invention, the negatively charged non-polymeric nanoparticles are silicon nanoparticles wherein the silicon nanoparticles are free from any drugs other than the one or more immunogenic drugs causing the unwanted immune response and optionally one or more immune modulating drugs.

[0096] Solid lipid

[0097] In one aspect of the present invention, the non-polymeric nanoparticles are solid lipid particles comprising at least one immune modulating drug(s).

[0098] In a preferred aspect of the present invention, the solid lipid particles comprise a negatively charged substance, such as a fatty acid or a negatively charged phospholipid.

[0099] In a more preferred aspect of the present invention, the solid lipid particles comprise at least one negatively charged substance, such as fatty acids or negatively charged phospholipids, and at least one PEG-lipid conjugate.

[0100] In another more preferred aspect of the present invention, the solid lipid nanoparticles comprise at least one negatively charged substance, such as fatty acids or negatively charged phospholipids, and cholesterol.

[0101] In another more preferred aspect of the present invention, the solid lipid particles comprise a negatively charged substance and l,2-distearoyl-sn-glycero-3 -phosphorylcholine (DSPC).

[0102] In another preferred aspect of the present invention, the negatively charged non-polymeric nanoparticles are negatively charged lipid particles wherein the lipid nanoparticles are free from any drugs other than the at least one immune modulating drug(s). This aspect of the 91.168061 / 01 invention relates to plain lipid particles without any drug substances other than the at least one immune modulating drug(s).

[0103] In another preferred aspect of the present invention, the negatively charged non-polymeric nanoparticles are negatively charged lipid particles wherein the lipid nanoparticles are free from any drugs other than the immune modulating drug(s) and optionally the immunogenic drug(s). No additional drug substances are present.

[0104] Liposomes

[0105] In one aspect of the present invention, the negatively charged non-polymeric nanoparticles are liposomes comprising at least one immune modulating drug(s).

[0106] In a preferred aspect of the present invention, the one or more immune modulating drugs and optionally one or more immunogenic drugs are encapsulated within the liposomes. This has the potential to selectively deliver said drugs at environments with low pH. For example, the pH in the target cells (e.g. dendritic cells) is approximately 2 to 5, such as 3 to 4.

[0107] In another preferred aspect of the present invention, the liposomes comprise at least one negatively charged substance, such as fatty acids or negatively charged phospholipids. For example, negatively charged phospholipids may include l-myristoyl-2-hydroxy-sn-glycero-3- phosphatidylethanolamine (PE), l-palmitoyl-2-hydroxy-sn-glycero-3-PE, l-oleoyl-2- hydroxy-sn-glycero-3-PE, l,2-dilauroyl-sn-glycero-3-PE, l,2-Dimyristoyl-sn-glycero-3-PE,

[0108] 1.2-Dipalmitoyl-sn-glycero-3-PE, 1,3 -dipalmitoyl glycero-2-PE, 1,2-diphytanoyl-sn-glycero- 3-PE and (DOPE (l,2-dioleoyl-sn-glycero-3-phosphoethanolamine) or phosphatidylcholines such as l-myristoyl-sn-glycero-3 -phosphatidylcholine, l-palmitoyl-sn-glycero-3- phosphocholine, 1 -stearoyl- sn-glycero-3 -phosphocholine, 1 -oleoyl-2-hy droxy- sn-glycero-3 - PC(phosphatidylcholine), rac-3-hexadecanamido-2-methoxypropan-l-ol phosphocholine, 2- arachidonoyl-sn-glycero-3 -phosphocholine, 1 ,2-dihexanoyl-sn-glycero-3 -PC, 1 ,2-dioctanoyl PC, l,2-didecanoyl-sn-glycero-3-phosphocholine, 1,2-dilauroyl- sn-glycero-3 -phosphocholine,

[0109] 1.2-dimyri stoyl- sn-glycero-3 -phosphocholine, 1 -myri stoyl-2-palmitoyl- sn-glycero-3 - phosphocholine, 1 -myri stoyl-2-stearoyl- sn-glycero-3 -phosphocholine, 1 -Palmitoyl-2-lauroyl- sn-gly cero-3 -PC, 1 -palmitoyl-2-myri stoyl- sn-glycero-3 -phosphocholine, 1 ,2-dipalmitoyl-sn- glycero-3 -phosphatidylcholine, l-palmitoyl-2-stearoyl-sn-glycero-3-phosphocholine, 1,2- 91.168061 / 01 diheptadecanoyl-sn-glycero-3-PC, l-stearoyl-2-myristoyl-sn-glycero-3-phosphocholine, 1- stearoyl-2-myristoyl-sn-glycero-3-phosphocholine, DSPC (distearoylphosphatidylcholine), l,2-dinonadecanoyl-sn-glycero-3-PC, l,2-diarachidoyl-sn-glycero-3-PC, 1,2-dihexadecyl-sn- glycero-3-PC, l,2-dipalmitoyl-sn-glycero-3-N,N-dimethyl-PE, l,2-diphytanoyl-sn-glycero-3- PC, DOPC (l,2-dioleoyl-sn-glycero-3 -phosphocholine) , 1,2-dierucoyl-sn-gly cero-3 - phosphocholine, l,2-dilinoleoyl-sn-glycero-3 -phosphatidylcholine, l-palmitoyl-2-oleoyl-sn- glycero-3 -phosphocholine, l-stearoyl-2-oleoyl-sn-glycero-3 -phosphocholine, 16:0-23:2 diyne PC, l-stearoyl-2-docosahexaenoyl-sn-glycero-3-phosphocholine, 1,2-didocosahexaenoyl-sn- glycero-3-PC, 23:2 diyne PC [DC(8,9)PC] and 23:2 diyne PE [DC(8,9)PE] or phosphoglycerols such as l-myristoyl-2-hydroxy-sn-glycero-3 -PG (phosphatidylglycerol), 1- palmitoyl-2-hydroxy-sn-glycero-3-PG, l-stearoyl-2-hydroxy-sn-glycero-3-PG, 1,2-dilauroyl- sn-glycero-3-phosphorylglycerol, l,2-dimyristoyl-sn-glycero-3 -phosphoglycerol, 1,2- dipalmitoyl-sn-glycero-3-phospho-(l'-rac-glycerol), 1,2-distearoyl-sn-gly cero-3 - phosphogly cerol, 1 -oleoyl-2-hy droxy- sn-gly cero-3 -PG, 1 ,2-dioleoyl- sn-gly cero-3 - phosphoglycerol and l-palmitoyl-2-oleoyl-sn-glycero-3 -phosphoglycerol or phosphoserines such as 1,2-dilauroyl-sn-gly cero-3 -phospho-L-serine, 1,2-dimyristoyl- sn-gly cero-3 -phospho- L-serine, 1,2-dipalmitoyl-sn-gly cero-3 -phospho-L-serine, l,2-distearoyl-sn-glycero-3- phospho-L-serine, 2-oleoyl-l-palmitoyl-sn-gly cero-3 -phospho-L-serine or 1,2- dioctadecenoyl- sn-gly cero-3 -phosphoserine or PEG (polyethylene glycol)derivatives such as m-PEG-DMPE, m-PEG-DOPE (l,2-dioleoyl-sn-glycero-3 -phosphoethanolamine) and DSPE(l,2-distearoyl-sn-glycero-3-phosphorylethanolamine) -PEG-OH, other related compounds like DSPE-glutaric acid and DSPE-succinic acid.

[0110] Preferred phospholipids include negatively charged phosphatidyl serine (PS) phospholipids and PEGylated phospholipids.

[0111] In another preferred aspect of the present invention, the negatively charged non-polymeric nanoparticles are negatively charged liposomes wherein the liposomes are free from any drugs other than immune modulating drug(s).

[0112] In another preferred aspect of the present invention, the negatively charged non-polymeric nanoparticles are negatively charged liposomes wherein the liposomes are free from any drugs other than the one or more immune modulating drugs and optionally immunogenic drugs causing the unwanted immune response. No additional drug substances are present. 91.168061 / 01

[0113] Immunogenic drugs

[0114] In some aspects of the present invention, the one or more immunogenic drugs causing the one or more unwanted immune responses are protein drug substances, protein conjugates, nucleic acid-based drug substances or particulate (water-insoluble) drug formulations.

[0115] In another preferred aspect of the present invention, the protein drug substances causing the one or more unwanted immune responses are monoclonal antibodies. Examples of antibodies include bagovomab, abciximab, actoxumab, adalimumab, adecatumumab, aducanumab, afelimomab, afutuzumab, alacizumab pegol, alemtuzumab, alirocumab, altumomab pentetate, amatuximab, anatumomab mafenatox, anifrolumab, anrukinzumab, apolizumab, arcitumomab, aselizumab, atinumab, atlizumab, atorolimumab, bapineuzumab, basiliximab, bavituximab, bectumomab, belimumab, benralizumab, bertilimumab, besilesomab, bevacizumab, bezlotoxumab, bimagrumab, bivatuzumab mertansine, blinatumomab, blosozumab, brentuximab vedotin, briakinumab, brodalumab, canakinumab, cantuzumab mertansine, cantuzumab ravtansine, caplacizumab, capromab pendetide, carlumab, catumaxomab, cedelizumab, certolizumab pegol, cetuximab, citatuzumab bogatox, cixutumumab, clazakizumab, clenoliximab, clivatuzumab tetraxetan, conatumumab, concizumab, crenezumab, dacetuzumab, daclizumab, dalotuzumab, daratumumab, demcizumab, denosumab, detumomab, dorlimomab aritox, drozitumab, duligotumab, dusigitumab, ecromeximab, eculizumab, edobacomab, falizumab, efungumab, eldelumab, elotuzumab, elsilimomab, enavatuzumab, enlimomab pegol, enokizumab, enoticumab, ensituximab, epitumomab cituxetan, epratuzumab, erlizumab, ertumaxomab, etaracizumab, abegrinmab, etrolizumab, evolocumab, exbivirumab, faralimomab, farletuzumab, fasinumab, felvizumab, fezakinumab ficlatuzumab, figitumumab, flanvotumab, fontolizumab, foralumab, foravirumab, fresolimumab, fulranumab, futuximab, galiximab, ganitumab, gantenerumab, gavilimomab, gemtuzumab, gevokizumab, girentuximab, glembatumumab vedotin, golimumab, gomiliximab, guselkumab, ibalizumab, ibritumomab tiuxetan, icrucumab, igovomab, imciromab, imgatuzumab, inclacumab, indatuximab, iravtansine, infliximab, intetumumab, inolimomab, minotuzumab ozogamicin, ipilimumab, iratumumab, itolizumab, ixekizumab, keliximab, labetuzumab, lambrolizumab, lampalizumab, lebrikizumab, lemalesomab, lerdelimumab, lexatumumab, libivirumab, ligelizumab, lintuzumab, lirilumab, lodelcizumab, lorvotuzumab mertansine, lucatumumab, lumiliximab, mapatumumab, 91.168061 / 01 margetuximab, maslimomab, mavrilimumab, matuzumab, mepolizumab, metelimumab, milatuzumab, minretumomab, mitumomab, mogamulizumab, morolimumab, motavizumab, moxetumomab pasudotox, muromonab-CD3, nacolomab tafenatox, namilumab, naptumomab, estafenatox, narnatumab, natalizumab, nebacumab, necitumumab, nerelimomab, nesvacumab, nimotuzumab, nivolumab, bofetumomab merpentan, ocaratuzumab, ocrelizumab, odulimomab, ofatumumab, olaratumab, olokizumab, omalizumab, onartuzumab, ontuxizumab, oportuzumab monatox, oregovomab, orticumab, otelixizumab, otlertuzumab, oxelumab, ozanezumab, ozoralizumab, pagibaximab, palivizumab, panitumumab, pankomab, panobacumab, parsatuzumab, pascolizumab, pateclizumab, patritumab, pemtumomab, perakizumab, pertuzumab, pexelizumab, pidilizumab, pinatuzumab vedotin, pintumomab, placulumab, polatuzumab vedotin, ponezumab, priliximab, pritoxaximab, pritumumab, quilizumab, racotumomab, radretumab, rafivirumab, ramucirumab, ranibizumab, raxibacumab, regavirumab, reslizumab, rilotumumab, rituximab, robatumumab, roledumab, romosozumab, rontalizumab, rovelizumab, ruplizumab, samalizumab, sarilumab, satumomab pendetide, secukinumab, seribantumab, setoxaximab, sevirumab, sibrotuzumab, sifalimumab, siltuximab, simtuzumab, siplizumab, sirukumab, solanezumab, solitomab, sonepcizumab, sontuzumab, stamulumab, sulesomab, suvizumab, tabalumab, tacatuzumab tetraxetan, tadocizumab, talizumab, tanezumab, taplitumomab paptox, tefibazumab, telimomab aritox, tenatumomab, teneliximab, teplizumab, teprotumumab, ticilimumab tildrakizumab, tigatuzumab, tocilizumab, toralizumab, tositumomab, tovetumab, tralokinumab, trastuzumab, tregalizumab, tremelimumab, tucotuzumab, celmoleukin, tuvirumab, ublituximab, urelumab, urtoxazumab, ustekinumab, vantictumab, vapaliximab, vatelizumab, vedolizumab, veltuzumab, vepalimomab, vesencumab, visilizumab, volociximab, vorsetuzumab mafodotin, votumumab, zalutumumab, zanolimumab, zatuximab, ziralimumab, zolimomab aritox.

[0116] In another preferred aspect of the present invention, the one or more immunogenic drugs causing the one or more unwanted immune responses is an antibody-drug conjugate. Antibody-drug conjugates may include any antibody-drug conjugate or fragments or derivatives thereof. Examples of antibody-drug conjugates include trastuzumab emtansine, gemtuzumab ozogamicin, brentuximab vedotin, T-DM 1, inotuzumab ozogamicin, pinatuzumab vedotin, RG-7596, lifastuzumab vedotin, glembatumumab vedotin, coltuximab ravtansine (SAR-3419), lorvotuzumab mertansine (IMGN-901), indatuximab ravtansine (BT- 91.168061 / 01

[0117] 062), anti-PSMA ADC, labetuzumab-SN-38, MLN-0264, ABT-414, and milatuzumab doxorubicin.

[0118] In another aspect of the present invention, the one or more immunogenic drugs causing the one or more unwanted immune responses are nucleic acid-based drug substances for use in gene therapy, preferably DNA nucleic acid-based drug substances for use in gene therapy or RNA nucleic acid-based drug substances for use in gene therapy or nucleotides conjugated with other substances eg. lipids or sugar moieties

[0119] In another aspect of the present invention, the one or more immunogenic drugs causing the one or more unwanted immune responses are nucleic acid-based drug substance for use in gene therapy encapsulated into a virus vector.

[0120] In a preferred aspect of the present invention, the one or more immunogenic drugs causing the one or more unwanted immune responses are nucleic acid-based drug substances for use in gene therapy encapsulated into an adenovirus associated virus vector type 9 (AAV9).

[0121] In another aspect of the present invention, the one or more immunogenic drugs causing the one or more unwanted immune responses are in the form of a water-insoluble particulate. Examples of water-insoluble formulations approved by regulatory bodies include daunorubicin, cytarabine, irinotecan and amphotericin B (See A.A.H. Abdellatif et al. in Nanotechnology Review 2021, 10, 1941-1977).

[0122] In another aspect of the present invention, the one or more immunogenic drugs causing the one or more unwanted immune responses are drugs for treatment of cancer, autoimmune diseases, respiratory diseases, gastrointestinal diseases, cystic fibrosis, cardiovascular disease, central nervous system diseases, diabetes, hemophilia, AIDS and eye diseases. Preferable drugs in this aspect of the invention include for example nivolumab, trastuzumab, elotuzumab, dinutuximab, catumaxomab, tositumomab. brentuximab vedotin, aldesleukin, broadly neutralising antibodies (bNAbs) and infliximab.

[0123] Immune modulating drugs 91.168061 / 01

[0124] In one aspect of the present invention, which may be combined with any of the above aspects in which one or more immune modulating drugs may be present, the nanoparticles may comprise one or more immune modulating drugs.

[0125] In some embodiments, such drugs are selected from ATC group L04.

[0126] For example, the one or more immune modulating drug substances may be selected from the following regulatory approved drug substances or groups of drug substances. The L04 code listed before the drug name is the international ATC code for the specific compound.

[0127] Group L04AA selective immune modulators such as L04AA02 muromonab-CD3, L04AA03 antilymphocyte immunoglobulin (horse), L04AA04 antithymocyte immunoglobulin (rabbit), L04AA06 mycophenolic acid as my cophenolate mofetil, L04AA10 rapamycin (sirolimus), L04AA13 leflunomide, L04AA15 alefacept, L04AA18 everolimus, L04AA19 gusperimus, L04AA21 efalizumab, L04AA22 abetimus, L04AA23 natalizumab, L04AA24 abatacept, L04AA25 eculizumab, L04AA26 belimumab, L04AA27 fmgolimod, L04AA28 belatacept, L04AA29 tofacitinib, L04AA31 teriflunomide , L04AA32 apremilast, L04AA33 vedolizumab, L04AA34 alemtuzumab, L04AA35 begelomab, L04AA36 ocrelizumab, L04AA37 baricitinib, L04AA38 ozanimod, L04AA39 emapalumab, L04AA40 cladribine, L04AA41 imlifidase, L04AA42 siponimod, L04AA43 ravulizumab, L04AA44 upadacitinib, L04AA45 filgotinib, L04AA46 itacitinib, L04AA47 inebilizumab, L04AA48 belumosudil, L04AA49 peficitinib, L04AA50 ponesimod, L04AA51 anifrolumab, L04AA52 ofatumumab, L04AA53 teprotumumab, L04AA54 pegcetacoplan, L04AA55 sutimlimab, L04AA56 deucravacitinib, L04AA57 ublituximab, L04AA58 efgartigimod alfa, L04AA59 avacopan.

[0128] Group L04AB tumor necrosis factor alpha (TNF-a) inhibitors such as L04AB01 etanercept, L04AB02 infliximab, L04AB03 afelimomab, L04AB04 adalimumab, L04AB05 certolizumab pegol, L04AB06 golimumab, L04AB07 opinercept,

[0129] Group L04AC interleukin inhibitors such as L04AC01 daclizumab, L04AC02 basiliximab, L04AC03 anakinra, L04AC04 rilonacept, L04AC05 ustekinumab, L04AC07 tocilizumab, L04AC08 canakinumab, L04AC09 briakinumab, L04AC10 secukinumab, L04AC11 siltuximab, L04AC12 brodalumab, L04AC13 ixekizumab, L04AC14 sarilumab, L04AC15 sirukumab, L04AC16 guselkumab, L04AC17 tildrakizumab, L04AC18 risankizumab, L04AC19 satralizumab, L04AC20 netakimab, L04AC21 bimekizumab, L04AC22 spesolimab, L04AC23 olokizumab. 91.168061 / 01

[0130] Group L04AD calcineurin inhibitors such as L04AD01 ciclosporin, L04AD02 tacrolimus, L04AD03 voclosporin.

[0131] Group L04AX other immunomodulators such as L04AX01 azathioprine, L04AX02 thalidomide, L04AX03 methotrexate, L04AX04 lenalidomide, L04AX05 pirfenidone, L04AX06 pomalidomide, L04AX07 dimethyl fumarate, L04AX08 darvadstrocel, L04AX09 diroximel fumarate.

[0132] The one or more immune modulating drug substances may be any combination of the above.

[0133] In another preferred aspect of the invention, the one or more immune modulating drug substances are selected from inhibitors of mTOR. mTOR (mammalianTarget Of Rapamycin) is a protein kinase enzyme in humans and animals which is critical for cell growth. The target has obtained the name based on the historical discovery of the target based on rapamycin which was known to have antifungal effects, however. mTOR inhibitors are generally administered at high concentrations and frequencies to prevent transplant rejection and graft versus host disease. Under these dosage conditions these compounds behave as potent immunosuppressors, and might increase infection susceptibility and cause some adverse effects like stomatitis. These adverse effects would not be acceptable for the prevention of anti-drug reactions. However, mTOR inhibitors may also be administered at lower concentrations to preferentially target specialized compartments of the immune system and provide selective immune modulating function. The drug substances rapamycin (also known as sirolimus) and everolimus discussed above are both mTOR inhibitors. 91.168061 / 01

[0134] Rapamycin (Sirolimus) Everolimus

[0135] Several mTOR inhibitors are well known in the relevant field. Such well known mTOR inhibitors include, among others, temsirolimus (CCI-779).

[0136] Temsirolimus

[0137] These compounds are all macrocyclic lactones and are known to be hydrolytically unstable; especially under neutral and basic conditions. In addition, the compounds have very low aqueous solubility.

[0138] A particularly preferred group of immune modulating drugs in the context of the present invention is rapamycin and derivatives thereof, especially rapamycin.

[0139] Rapamycin derivatives described in the literature can, for example, be found in the following documents: WO2022206796, WO2023031738, WO2022159976, W02022216900,

[0140] WO2021133509, WO2021113665, WO2021086946 , W02021000899, WO2020249652, W02020194209, WO2020128861, WO2019241789, WO2019235879, W02020076728, W02019209083, WO2019157363, WO2019157382, WO2019157374, WO2019141254, WO2019126374, WO2019212990, WO2019115640, WO2019122065, WO2019122059, WO2019064182, WO2018187414, WO2018148508, WO2018204416, WO2017193562, WO2017083823, W02017044720, W02017040341, W02017011363, WO2017031427, W02016207205, W02016010869, WO2015192761, WO2015184983, W02015096640, WO2014154026, WO2014118696, WO2014116611, W02015004455, WO2014169167, WO2014022879, WO2014022728, WO2013182503, WO2013071698, W02013111106, W02013016164, W02013016160, WO2013093493, WO2012167606, W02013050508, W02013050419, WO2013041652, W02012047570, WO2012047569, WO2012136622, W02012131019, W02012027240, WO2012027239, WO2012027236, WO2012027234, 91.168061 / 01

[0141] W02012097039, W02012071519, W02012071509, WO2012074869, W02012071501, W02012071511, W02012068106, W02012068096, WO2011146594, WO2012017449, WO201 1123524, WO2011090935, WO2011080568, WO2011151704, WO2011141713, WO201 1107585, WO2011051960, W02010120994, WO2011029027, WO2011025889, WO201 1002887, WO2010138487, WO2010135568, W02010138490, WO2010126895, W02010056320, W02010120998, W02010118207, W02010120996, W02010114494, W02010120991, W02010120987, W02010044885, W02010031251, W02010103094, W02010096314, W02010039740, W02009131631, W02010030727, W02010030967, WO2009143313, W02010008847, W02010011620, W02010002954, W02009155052, W02009155042, WO2009122176, W02009070524, W02009117482, W02009111547, W02009097515, W02009097490, W02009052145, W02009091788, W02008148074, W02008124026, W02007087395, W02007067560, W02007061737, W02006096325, W02006068932, W02006068905, W02006039237, W02005105811, W02005100366, W02005047295, W02005023254, W02005105812, WO9915530, W09809970, WO9809972, WO9617845, WO9616967, WO9534565, WO9528406, WO9518133, WO95 14697, WO9514696, WO9514023, WO9504738, WO9522972, WO9425468, WO9425072, WO9425022, WO9412655, WO9410843, WO9410176, WO9411380, WO9402137, WO9402485, WO9402136, WO9318043, WO9310122, W09305046, WO9221341, WO9205179, W02004060283, W02004011000, W003018573, W00224706, W00151049, WO0134816, WO0123395, W02008065887, W02008032091, W02008032089, W02008032086, W02008032072, W02008032060, W02008032036, W02008032033, W02008032028, W02008023161, W02007135411, WO2007135397, W02007060404, W02006095185 and W02006090169.

[0142] In another preferred aspect of the invention, the one or more immune modulating drug substances are selected from specific compounds understood to have activity as inhibitors of mTOR. Some of these compounds are: A-443654, ABI009, AEZS-126, AEZS-127, AR-12, AR-42, AT13148, apitolisib, AZD6482, AZD8055, BGT226, BEZ235, bimiralisib, biolimus a9, CAL-101, CAL-120, CAL-263, CC-11, CC-223, GCS-0941, dactolisib, DB12180, everolimus, EXEL-2044, EXEL-3885, EXEL-4431, EXEL-7518, EZN-4150, farnesyl thioslicyclic acid, GDC-0369, GDC-0941, gedatolisib, GSK60-693, GSK1059615, GSK1720070, INK128, Ku-0063794, LY294002, LY3023414, LY317615, MHY1485, miltefosine, MK2206, MKC-1, KRX-0401, KU-006374, myolimus, novolimus, NV- 128, 91.168061 / 01

[0143] NVP-BEZ235, omipalisib, OSI-027, P2281, palmoid 529, PF-04691502, PI-103, PP121, PP242, PP30, PQR309 PX-866, QLT-0447, ridaforolimus, sapanisertib, SF-1126, SF2626, sirolimus (rapamycin), SR13668, TAE226, TAFA-93, temsirolimus, Torin 1, Torin 2, TOP216, vistusertib, voxtalisib, VQD-002, WAY-600, WJD008, wortmannin, WX-037, WYE-354, WYE-687, WYE-125132, XL147, XL388, XL-765 YM-58483, zotarolimus and 3BDO.

[0144] Composition formulations, route of administration dosing and dose regime

[0145] There are several formulation options depending upon the clinical situation and choice of drug substances.

[0146] Option 1 - A single composition comprising negatively charged non-polymeric nanoparticles free from any drug (plain nanoparticles) and one or more immunogenic drugs not associated with the nanoparticles in the same formulation. One administration.

[0147] Option 2 - Two separate compositions, wherein composition 1 comprises nanoparticles free from any drug (plain nanoparticles) and composition 2 comprises one or more immunogenic drugs including fragments of such drugs. The two compositions may be mixed prior to administration and administered together or administered separately. Composition 1 may be administered prior to, simultaneously with or after composition 2. When administered separately, there may be a time lag between the administrations. This time lag may be 1 to 60 minutes, 1 to 24 hours, 1 to 10 days or any combination thereof. The most appropriate time periods will vary depending upon choice of drugs and the clinical indication. Typical time lag can be within less than 24 hours.

[0148] The product may be a kit comprising two separate formulations: the drug free particles and the one or more macromolecular drugs. Another option is that the one or more immunogenic drugs are a separate commercial product and not a part of a kit. If the product is in two separate formulations, the immunogenic product may be from two different producers with two different regulatory approvals. 91.168061 / 01

[0149] Option 3 - One composition comprising the negatively charged non-polymeric nanoparticles and one or more immunogenic drugs or fragments of such drugs, wherein the one or more immunogenic drugs are encapsulated in the negatively charged non-polymeric nanoparticles.

[0150] In this scenario, the nanoparticles have two potential functions:

[0151] 1. Delivery of the immunogenic drug; and

[0152] 2. The negatively charged non-polymeric nanoparticles as such prevent or reduce unwanted immune response from the immunogenic drug.

[0153] Option 4 - One composition comprising the negatively charged non-polymeric nanoparticles, one or more immune modulating drug substances and one or more immunogenic drugs, wherein the one or more immune modulating drug substances are encapsulated in the negatively charged non-polymeric nanoparticles. The one or more immunogenic drugs are typically dissolved in an aqueous phase in the same composition.

[0154] Option 5 - Two compositions. One composition comprises the negatively charged non- polymeric nanoparticles and one or more immune modulating drug substances, wherein the one or more immune modulating drug substances are encapsulated in the negatively charged non-polymeric nanoparticles. Another composition comprises the one or more immunogenic drugs. The two compositions may be mixed prior to administration and administered together or administered separately. One may be administered prior to, simultaneously with or after the other composition. When administered separately, there may be a time lag between the administrations. This time lag may be 1 to 60 minutes, 1 to 24 hours, 1 to 10 days or any combination thereof. The most appropriate time periods will vary depending upon choice of drugs and the clinical indication. Typical time lag can be within less than 24 hours.

[0155] The product might be a kit comprising two separate formulations: (1) a formulation comprising the immunogenic drug free particles and (2) a formulation comprising the macromolecular drug. Another option is that the one or more immunogenic drugs are a separate commercial product and not a part of a kit. If it is in two separate formulations, the 91.168061 / 01 immunogenic product may be from two different producers with two different regulatory approvals.

[0156] All options have the potential to prevent or reduce the unwanted immune responses generated by use of the immunogenic drugs and thereby improve the efficacy and safety of the clinical use said drugs compared to use without the nanoparticulate material. The present nanoparticulate compositions can improve unwanted immune responses to a level where the patients do not have to stop or change treatment regime over time.

[0157] The present nanoparticulate compositions can allow for the potential use of new immunogenic drugs that have clinical potential but have so far not been used clinically due to the resultant unwanted immune responses. Therefore, immunogenic drugs that may previously have been thought unsafe have potential for future use in combination with the present nanoparticulate compositions.

[0158] The compositions according to the present invention can be administered locally or systemically. A preferred administration route is parenteral administration. The preferred form for administration is by injection, typically intravascular injection, intravenous injection intramuscular injection, subcutaneous injection or intranasal administration. More preferably, administration is by intravenous injections or subcutaneous injections. If possible, a subcutaneous injection may be preferred.

[0159] The preferred administration route depends on the indication, the choice of drug substances and where the treatment is to take place.

[0160] Both the particulate compositions and the compositions comprising the one or more immunogenic drug substances can be in the form of ready to use or in the form of a dry material to be dissolved and / or suspended and / or diluted before use.

[0161] Preferred dry compositions of the present nanoparticles are compositions where the nanoparticulate material has been dried by freeze drying or by drying in fluid bed. Other gentle methods to secure minimal degradation and easy suspension of free nanoparticles by adding water before administration are also contemplated. 91.168061 / 01

[0162] In some aspects, the most preferred formulations comprising the composition for use according to the present invention do not need any ultrasound treatment nor any other suspension / dissolution method prior to use.

[0163] The formulations may optionally be administered with a filter in the line before administration to the subject. This is to ensure that larger particle aggregates (much larger than 500 nanometers) are not administered to the subject. The use of a filter to eliminate potential particle aggregates is of particular use when particulate drugs are injected; especially for intravenous injections.

[0164] In some aspects, the most preferred formulations comprising the composition for use according to the present invention do not need any form of filter since the suspension / dissolution method ensures adequate dispersion of the nanoparticles.

[0165] In one aspect, the compositions can be in any pharmaceutical container; typically, for example, injection vials of glass with a septum, vials of polypropylene or prefilled syringes. The injection can take place by classical hand-driven injection or by use of an injection device.

[0166] In one aspect, the final compositions for administration may be isotonic or weak hypertonic. The preferred osmolality (concentration in blood) may be below 900 mOsm / kg, more preferably below 600 mOsm / kg, most preferably around 300 mOsm / kg. Osmolality can be determined by using commercially available osmometers.

[0167] In one aspect, the compositions are aqueous compositions, optionally comprising small amounts of non-toxic water-soluble organic solvents. Preferably, the compositions according to the present invention do not comprise organic solvents.

[0168] Both the nanoparticulate compositions and compositions comprising one or more immunogenic drugs may, according to the present invention, typically comprise various pharmaceutical excipients. These excipients can include typical stabilizing excipients, pH- adjusting agents, osmolality regulating compounds and antioxidants. Typical excipients include for example pH-adjusting agents like HC1 and NaOH, buffer substances like acetate, phosphate, histidine, and tris (tromethamin), osmolality regulating compounds like glucose, sucrose and trehalose and antioxidants like chelating agents (e.g. EDTA) and sodium metabisulfite. 1 91.168061 / 01

[0169] The pH in the final formulations for administration may be around 7.4. In some cases, it may, for stability purposes, be necessary to have compositions with a lower pH. This is generally acceptable if the injection volume is relatively low. The pH might be as low as 3 if the injection volume is 1 ml or less. Preferred formulations have however pH above 5.

[0170] Osmolality regulating compounds can be pharmaceutical acceptable salts or non-ionic compounds such as sugars. Experiments with the present nanoparticles have shown that suspensions of the present nanoparticles are more stable if salts are not present or present in low concentrations. Preferred osmolality regulating additives in the present compositions comprising nanoparticles may be neutral agents such as glucose, sucrose, trehalose or any combination thereof.

[0171] In one aspect, at least one of, preferably both, the nanoparticulate compositions and the compositions comprising one or more immunogenic drugs may include one or more surface active excipients for stabilization of the nanoparticles and optionally solubilization of potential immunogenic drug substances. Any pharmaceutically acceptable surfactant might be used. Preferable surfactants are surfactants with relatively high hydrophilic-lipophilic balance value HLB value, for example polysorbate 80 and / or polysorbate 40 with HLB- value around 15 on the scale from 0 to 20.

[0172] In one aspect, the nanoparticulate material according to the present invention may be coated by non-covalent adsorption of a surface active material or covalently coated by chemical compounds for the modification of particle surface and / or to modify the release of the one or more drug substances from the particles.

[0173] For example, silicon particles can be coated non-covalently with proteins such as albumin, surfactants and / or fat derivatives.

[0174] Silicon particles according to one aspect of the present invention can be covalently modified by forming silicon-oxygen-silicon bonds using silane reagents, for example as described in M. P. Schwartz, F. Cunin, R. ffl. Cheung and M. J. Sailor, Chemical modification of silicon surfaces for biological applications, Phys. Status Solidi A, 2005, 202(8), 1380 1384. Silicon particles according to one aspect of the present invention can also be covalently surface modified by formation of silicon-carbon bonds; for example, by reaction with an unsaturated fatty acid forming a carboxylated particle or by reaction with a polyethylene derivative forming a surface with increased hydrophilicity. 91.168061 / 01

[0175] The immune modulating drugs to be incorporated into the nanoparticles can, according to one aspect of the present invention, be in the form of a pharmaceutically acceptable salt or complex with a pharmaceutically acceptable compound. Preferred pharmaceutically acceptable salts for immune modulating drug substances with an acidic group include, for example, sodium salts, potassium salts and meglumine salts. Preferred pharmaceutically acceptable salts for immune modulating drug substances with a basic group include, for example, chloride salts, oxalate salts and mesylate salts. The choice of salt depends on the selection of the one or more immune modulating drug substances and factors such as stability and water solubility.

[0176] One preferred form of the one or more immune modulating drug substances within the nanoparticle is in the form of a complex with cyclodextrins. The most preferred cyclodextrins are beta-cyclodextrin, 2-hydroxypropyl-beta-cyclodextrin and 4-sulphobutyl-beta- cyclodextrin. Typical examples are rapamycin 2-hydroxypropylyl-beta-cyclodextrin and rapamycin 4-sulphobutyl-beta-cyclodextrin. Alpha- and gamma-cyclodextrin derivatives may also be used for other immune modulating agents.

[0177] Several of the immune modulating drug substances have high affinity to proteins. Rapamycin has, in a clinical situation, a very high degree of albumin binding. For example,

[0178] WO202 1086946 describes rapamycin albumin nanoparticles for the treatment of cancer. One option of the present invention relates to use of immune modulating drugs in the form of albumin complexes in the present negatively charged non-polymeric nanoparticles.

[0179] A preferred option according to one aspect of the present invention is the use of the immune modulating drugs in the form of complexes with the macromolecular agent within the nanoparticles. A typical example is rapamycin trastuzumab complexes. Complexes between proteins and immune modulating agents such as, for example, rapamycin is a non-covalent complex which in vivo, like other drug-protein complexes, will dissociate to the immune modulating drug and the macromolecular drug substance.

[0180] Several of the immune modulating drugs which may be incorporated into the present nanoparticles are unstable compounds with low aqueous solubility. The present salts and complexes have the potential to improve both solubility and stability of the immune modulating drug substances. The inventors have, for example, shown that rapamycin, when 91.168061 / 01 complexed with 2-hydroxypropyl-beta-cyclodextrin or 4-sulphobutyl-beta-cyclodextrin has a much higher aqueous solubility than rapamycin alone.

[0181] Stabilizing excipients are typically buffers such as, for example, histidine buffers. Other potential additives include, for example, viscosity adjusting excipients and antioxidants.

[0182] The preferred compositions according to the present invention are sterile compositions.

[0183] Sterile compositions comprising nanoparticles and / or drug substances according to the present invention can be prepared by aseptic production technology and / or final sterilization methods well known in the art.

[0184] The dosage and dosing protocol for the immunogenic drugs according to the present invention may be comparable or about the same as the current dosage or dosing protocol for the approved immunogenic drug. Clinical experience in the future may allow for an increase in the dosage or an increase in the frequency of dosing based on the immune modulating effect of the nanoparticles of the present invention.

[0185] The dosage and dosing schedule for the nanoparticulate composition may vary dependent on the clinical situation, choice of macromolecular drug substance, presence or lack thereof of immune modulating drugs within the nanoparticles and the choice of one or more immune modulating drugs.

[0186] The dose of immune modulating drug may vary from about 0 to 300 mg per day. The dose varies depending upon choice of immune modulating drug and the clinical indication. For rapamycin and everolimus the calculated typical daily dose over time is between 0.01 and 20 mg.

[0187] The amount of nanoparticulate material may vary from 0.01 mg to 200 mg per day. The dose varies depending upon choice of immune modulating drug and the clinical indication. For nanoparticles comprising rapamycin or everolimus the calculated typical daily dose of nanoparticles over time is between 0.1 and 100 mg.

[0188] The dosing frequency may vary from once daily dosing to weekly dosing or less frequent. Typical dosing frequency could be once weekly, every second week or every 4 week. The dosing frequency is depending upon choice of immune modulating drug and the clinical indication. The more frequent the ADA forming drug is dosed, the more frequently is the present nanoparticulate drug dosed. 91.168061 / 01

[0189] Brief description of Figures:

[0190] Figure 1. ADA responses following administration of the immunogenic drug candidate NJH395 and reduction of these responses when administered with rapamycin (rap) and rapamycin-HP-P-CD loaded (rap:CD) cCVD Si (Sil) particle formulations. Figure 1 (a) shows the IgG titers and Figure 1 (b) shows the IgM titers as the average of 8 datapoints with error bars showing standard error of the mean (SEM). This figure shows the data produced from the experiment in example 2.

[0191] Figure 2. ADA responses following administration of the immunogenic drug candidate NJH395 and reduction of these responses when administered with rapamycin-HP-P-CD loaded (rap:CD) cCVD Si (Sil) particle formulations. Figure 2 (a) shows the IgG titers and Figure 2 (b) shows the IgM titers as the average of 8 datapoints with error bars showing standard error of the mean (SEM). Figure 2 (c) shows the analyses of spleens from the groups administered with the rapamycin-containing formulation, with empty particles and with NJH395 only. This figure shows the data produced from the experiment in example 3.

[0192] Figure 3. ADA responses following administration of the immunogenic drug candidate NJH395 and reduction of these responses when administered with lyophilized rapamycin-HP- P-CD loaded (rap-CD) or empty DPPC-coated cCVD Si nanoparticle (SiNP) formulations. Figure 3 (a) shows the IgG titers and Figure 3 (b) shows the IgM titers as the average of 8 datapoints with error bars showing standard error of the mean (SEM). This figure shows the data produced from the experiment in example 6.

[0193] The present particles, compositions and their use in biological experiments are further illustrated by the non-limiting examples below.

[0194] Examples

[0195] Example 1: Preparation of porous silicon nanoparticles with or without rapamycin or cyclodextrin complexes of rapamycin 91.168061 / 01

[0196] Rapamycin-HP-P-CD complexes were made by mixing rapamycin and HP-P-CD (1 : 1.1, rapamycimCD mole ratio) with a small amount of water with an Eppendorf micro-pestle, before evaporating the water till dryness under vacuum. Synthesis of Si particles was done by centrifugal Chemical Vapor Deposition (cCVD) process. Empty, Pluronic-F127 treated cCVD Si particles (formulation 1.1) were prepared by addition of Pluronic-F127 solution to cCVD Si particles, ultrasonication treatment for 15 minutes followed by two water washing-steps after centrifugation, followed by vacuum drying. Rapamycin, or the prepared Rapamycin-HP- P-CD complexes, were loaded into Pluronic-F127 treated cCVD Si particles using the solvent evaporation technique. First, rapamycin, or rapamycin-HP-P-CD, was dissolved in dimethylformamide (DMF). Then, the particles were added to obtain three different particle formulations: 5wt% rapamycin loaded (formulation 1.2), 5wt% rapamycin with rapamycin- HP-P-CD loaded (formulation 1.3), and 10wt% rapamycin with rapamycin-HP-P-CD loaded particles (formulation 1.4). The samples were immersed in an ultrasonicator bath for 15 minutes before they were dried by evaporating the DMF under vacuum.

[0197] The empty control particles possessed a diameter of 200 nm, PDI of 0.19 and zeta potential of -38 mV. The prepared formulations had a particle size of 175 nm, PDI of 0.16 and zeta potential of -38 mV for rapamycin-loaded ones, and 180 nm, PDI of 0.18 and zeta potential of -38 mV for rapamycin-HP-P-CD loaded ones (5 wt% rapamycin), as measured in water using Dynamic Light Scattering.

[0198] Example 2: ADA efficacy study in mice over 13 days

[0199] All procedures involving animal experiments described in this document were performed under the authority of the Cantonal Veterinary Office of Basel city and meet the standards required by the Swiss BLV guidelines. The ADA experiments (example 2, 3, 4, 6 and 7) were conducted with C57 black 6 mice. The health status of the mice was continuously monitored over multiple weeks.

[0200] Both formulations prepared in example 1 with 5wt% rapamycin were tested for their ADA- mitigation abilities against the immunogenic drug candidate NJH395 in mice models. Two master-mixes of the 5% rapamycin-loaded particle formulations (with and without HP-P-CD complex) with NJH395 were prepared in PBS, before ultrasonication treatment to redisperse the particle dispersion. The formulations were then injected subcutaneously to female black 6 91.168061 / 01 mice. 8 weeks old animals, with 8 animals per group, were given either NJH395 (0.5 mg / kg) plus either rapamycin-loaded particles (formulation 1.2), rapamycin- HP-P-CD loaded particles (formulation 1.3), free rapamycin- HP-P-CD complex or empty particles (formulation 1.1). The rapamycin dose was 2.5 mg / kg for each animal and each formulation / group. Subcutaneous administration was done on day 0, 2, 7 and 9, and the animals were sacrificed on day 13. The anti-NJH395 IgM and IgG titers were measured by ELISA assays in blood samples taken on day 0, 7, 9 and 13.

[0201] The results showed that the rapamycin-HP-P-CD loaded particles gave the strongest reduction in ADA responses, comparable with rapamycin-HP-P-CD free, as measured by the levels of anti-NJH395 IgG and IgM antibody production (see Figure 1). On days 9 and 13, the average IgG titers from the Si loaded rapamycin-HP-P-CD group were reduced to about 5% and 1% of the corresponding titer levels from the empty Si particle formulation, respectively. The average IgM titers were reduced to 38% on day 7, while returning to similar levels on day 9, confirming a rapid response. The rapamycin-loaded Si (without CD complex) formulation reduced the IgG titers to 48% of empty particles on day 13.

[0202] Example 3: ADA efficacy study in mice over 32 days

[0203] The rapamycin-HP-P-CD loaded Si particle formulation prepared with 10wt% rapamycin in example 1 was tested for mitigation of anti-NJH395 antibodies in vivo in C57 black 6 mice. A master mix of NJH395 and rapamycin-HP-P-CD loaded Si particles was prepared in glucose (5%, isotonic) and treated in ultrasonicator bath for redispersion before injection. Three groups of female black 6 mice (8 animals / group, 8 weeks old) were given NJH395 alone (1 mg / kg) or with either rapamycin-HP-P-CD loaded particles (formulation 1.4, 1 mg / kg rapamycin dose) or empty particles (formulation 1.1, corresponding particle concentration) on day 0 and 5 (all formulations were given by subcutaneous administration). Keyhole Limpet Hemocyanin (KLH) was administered by intraperitoneal injection at 5 mg / kg to test specificity of the induced immune tolerance, on day 10 and 17. NJH395 alone was administered subcutaneously to all animals on day 17 for testing the duration of the response. After terminating the experiment on day 32, the mice were sacrificed. The spleens from two animals per group were sampled to analyze the amounts of CD25+ (APC-A) and FoxP3+ ( Alexa Fluor 488-A) T regulatory cells by flow cytometry. The anti-NJH395 and anti-KLH 91.168061 / 01

[0204] IgM and IgG titers were measured by ELISA assays in blood samples taken at days 0, 5, 10, 17, 24 and 32.

[0205] The results showed a reduction in both IgG and IgM endpoint titers for both the particle- treated groups, with a stronger response from the rapamycin-containing formulations (see Figure 2). IgM titers from the particle formulations were reduced to 28-38% of the NJH395 control on day 5 and 24, with similar levels at the other timepoints. This corresponds with a rapid and short-lived response following the administration pattern. IgG titers were comparable until days 24 and 32, where they were reduced to 8% and 12% with the rapamycin formulation and to 27% and 23% with the empty particles, respectively, compared to NJH395 control levels. Furthermore, anti-KLH-antibodies were not affected to the same extent. Anti-KLH IgG titers reduced to approximately 30% and 52% for the empty particles on days 24 and 32, respectively. At the other time-points, and for the rapamycin loaded particles, IgG and IgM titers remained mostly within ±15% of the KLH control titers. This indicates that the immune response induced by the RAP particles is specific to NJH395. Analyses of spleens from two of the animals per group showed an upregulation of T regulatory (CD3+CD4+CD25+FoxP3+) cells to 6.3-6.7% with the rapamycin-containing formulation, compared to 4.8-5.4% with empty particle treatment 3.6-3.8% with NJH395 only (Figure 2(c)). These data suggest that co-administration with the cCVD Si-rapamycin-HP-P- CD formulation can reduce ADA responses towards NJH395. Additionally, these data suggest that empty Sil particles also affect the immune responses, though less strongly than rapamycin-containing ones.

[0206] Example 4: Intravenous tolerability study of silicon nanoparticles

[0207] The pluronic-F127 treated cCVD Si particles (empty control particles) prepared in example 1 (formulation 1.1) were investigated for their tolerability after intravenous injection in C57 black 6 mice. The particle samples were dispersed in isotonic glucose solution before intravenous administration with single, ascending doses of 0.1, 1, 5 and 25 mg / kg particles, with each dose given to at least two animals. The highest dose (25 mg / kg) was administered to three animals. Blood samples were withdrawn before dosing and after 0.25, 1, 2, 4, 7 and 24 hours to examine hematological and clinical chemistry parameters. The animals were 91.168061 / 01 closely monitored for any side effects. Their bodyweight was measured at study start and at termination (24 hours).

[0208] The results showed that all dosed groups were tolerated (dose up to 25 mg / kg) with no clinical signs of toxicity at any time point, and the mice behaved normal. For all groups, 15 minutes after injection the glycemia values increased, as a result of the glucose content in the formulation, but at the Ih time point the baseline values were reached again. Bodyweight was moderately affected by all groups, but in an acceptable range in these settings, as also bleeding time points could cause stress and loss in bodyweight for the animals. There were no clinical pathology findings or no evidence of toxicity.

[0209] Example 5: Preparation of lyophilized formulation of porous silicon nanoparticles with rapamycin-HP-P-CD with a dipalmitoylphosphatidylcholine (DPPC) coating

[0210] DPPC treated control particles (formulation 5.1) were prepared by dissolving DPPC in toluene, adding the cCVD Si particles (prepared by centrifugal Chemical Vapor Deposition), placing the dispersion in an ultrasonicator bath for 15 minutes and drying under vacuum. Rapamycin-CD loaded particles (10 wt% rapamycin) were prepared by dissolving rapamycin, HP-P-CD and cCVD Si particles in a solution of 1 -propanol: water 1 : 1, sonication treatment of the dispersion for 30 minutes, followed by drying under vacuum. The loaded particles were coated with DPPC (formulation 5.2) by dissolving the two substances in toluene, sonication treatment for 15 minutes and solvent removal under vacuum. Lyophilization of formulation 5.1 and 5.2 was done, separately, by adding an isotonic sucrose solution, freezing the samples under liquid nitrogen and freeze drying overnight. The vials were then filled with argon and sealed for storage.

[0211] The empty DPPC-coated particles possessed a diameter of 288 nm, PDI of 0.27 and zeta potential of -30 mV. The prepared rapamycin-containing DPPC-coated formulation had a particle size of 289 nm, PDI of 0.24 and zeta potential of -32 mV, as measured in water using Dynamic Light Scattering.

[0212] Example 6: ADA efficacy study in mice over 24 days with lyophilized particle formulation 91.168061 / 01

[0213] The rapamycin-HP-P-CD loaded and DPPC coated Si particle formulation prepared in example 5 was tested for mitigation of anti-NJH395 antibodies in vivo with C57 black 6 mice (8 mice / group). NJH395 was administered subcutaneously with a dose of 1 mg / kg on day 0, 5 and 10, alone or together with redispersed particle formulation of either the empty control particles (formulation 5.1) or with rapamycin-load (formulation 5.2) with a dose of 25 mg / kg (2.5 mg / kg rapamycin dose). NJH395 (subcutaneously) and KLH (intraperitoneally, 5 mg / kg) were administered alone on day 17. The anti-NJH395 and anti-KLH IgM and IgG titers were measured by ELISA assays in blood samples taken at days 0, 5, 10, 17 and 24, and the mice were terminated on day 24.

[0214] The results showed that the rapamycin-HP-P-CD loaded particles reduced the anti-NJH395 IgG antibody titers to 9% and 12% of the titer levels for NJH395 administered alone on day 17 and 24, respectively. The similar comparison gave reduced anti-NJH395 IgM antibodies to 34% on day 5, compared to NJH395 alone. This shows that the rapamycin-HP-P-CD loaded DPPC-coated cCVD Si particle formulation can be useful for reducing ADA responses towards the biotherapeutic NJH395. The anti-KLH antibodies, on the other hand, showed higher titers for the rapamycin-loaded formulation on day 24, showing that the immune suppression is specific towards the biotherapeutic NJH395. The results are shown in Figure 3.

[0215] Example 7: Observation of safety of silicon nanoparticles during various efficacy studies in mice

[0216] Several mice (more than 150) have been studied in several rounds of anti- ADA efficacy studies administering various doses of formulations of empty cCVD silicon particles (formulation 1.1 and 5.1) and cCVD silicon particles comprising rapamycin (formulation 1.2) or rapamycin-cyclodextrin complexes (formulation 1.3, 1.4 and 5.2), or variations of these. The particle formulations were administered by subcutaneous injections, and the doses typically contained 10-25 mg / kg of particles, corresponding to 1-2.5 mg / kg of rapamycin. Each mouse was typically subject for 3-4 injections containing the particle formulations. The mice were closely observed over the study duration that varied between 12 and 56 days. No drug-related mortality was observed. The body weight, clinical, behavioral and histological findings showed no evidence of toxicity.

Claims

91. 168061 / 01Claims1. A composition for use in the treatment or prevention of one or more unwanted immune responses in a subject, wherein the one or more unwanted immune responses are caused by the use of one or more immunogenic drugs and wherein the composition comprises negatively charged non-polymeric nanoparticles, wherein the negatively charged non- polymeric nanoparticles are selected from the group consisting of at least one of silicon particles, solid lipid particles comprising at least one immune modulating drug and liposomes comprising at least one immune modulating drug.

2. The composition for use as claimed in claim 1, wherein the composition further comprises the one or more immunogenic drugs, or fragments thereof, optionally wherein the one or more immunogenic drugs or fragments thereof are encapsulated within the negatively charged non-polymeric nanoparticles.

3. The composition for use as claimed in claim 1 or claim 2, wherein the negatively charged non-polymeric nanoparticles are silicon particles and the composition further comprises one or more immune modulating drugs, where said immune modulating drug is not encapsulated within the negatively charged non-polymeric nanoparticles.

4. The composition for use as claimed in any of claims 1 to 3, wherein the unwanted immune response is the production of one or more anti-drug antibodies (AD As) or cytotoxic responses.

5. The composition for use as claimed in any of claims 1 to 4, wherein the one or more immunogenic drugs are selected from the group consisting of proteins and protein derivatives, preferably wherein the one or more immunogenic drugs are selected from at least one of monoclonal antibodies, monoclonal antibody conjugates or nucleic acids.

6. The composition for use as claimed in any of claims 1 to 5, wherein the one or more immune modulating drugs are selected from ATC group L04, preferably selected from: group L04AA selective immunomodulators such as L04AA02 muromonab-CD3, L04AA03 antilymphocyte immunoglobulin (horse), L04AA04 antithymocyte68061 / 01 immunoglobulin (rabbit), L04AA06 mycophenolic acid as mycophenolate mofetil, L04AA10 rapamycin (sirolimus), L04AA13 leflunomide, L04AA15 alefacept, L04AA18 everolimus, L04AA19 gusperimus, L04AA21 efalizumab, L04AA22 abetimus, L04AA23 natalizumab, L04AA24 abatacept, L04AA25 eculizumab, L04AA26 belimumab, L04AA27 fingolimod, L04AA28 belatacept, L04AA29 tofacitinib, L04AA31 teriflunomide, L04AA32 apremilast, L04AA33 vedolizumab, L04AA34 alemtuzumab, L04AA35 begelomab, L04AA36 ocrelizumab, L04AA37 baricitinib, L04AA38 ozanimod, L04AA39 emapalumab, L04AA40 cladribine, L04AA41 imlifidase, L04AA42 siponimod, L04AA43 ravulizumab, L04AA44 upadacitinib, L04AA45 filgotinib, L04AA46 itacitinib, L04AA47 inebilizumab, L04AA48 belumosudil, L04AA49 peficitinib, L04AA50 ponesimod, L04AA51 anifrolumab, L04AA52 ofatumumab, L04AA53 teprotumumab, L04AA54 pegcetacoplan, L04AA55 sutimlimab, L04AA56 deucravacitinib, L04AA57 ublituximab, L04AA58 efgartigimod alfa, L04AA59 avacopan; group L04AB tumor necrosis factor alpha (TNF-a) inhibitors such as L04AB01 etanercept, L04AB02 infliximab, L04AB03 afelimomab, L04AB04 adalimumab, L04AB05 certolizumab pegol, L04AB06 golimumab, L04AB07 opinercept; group L04AC interleukin inhibitors such as L04AC01 daclizumab, L04AC02 basiliximab, L04AC03 anakinra, L04AC04 rilonacept, L04AC05 ustekinumab, L04AC07 tocilizumab, L04AC08 canakinumab, L04AC09 briakinumab, L04AC10 secukinumab, L04AC11 siltuximab, L04AC12 brodalumab, L04AC13 ixekizumab, L04AC14 sarilumab, L04AC15 sirukumab, L04AC16 guselkumab, L04AC17 tildrakizumab, L04AC18 risankizumab, L04AC19 satralizumab, L04AC20 netakimab, L04AC21 bimekizumab, L04AC22 spesolimab, L04AC23 olokizumab; group L04AD calcineurin inhibitors such as L04AD01 ciclosporin, L04AD02 tacrolimus, L04AD03 voclosporin; and group L04AX other immunomodulators such as L04AX01 azathioprine, L04AX02 thalidomide, L04AX03 methotrexate, L04AX04 lenalidomide, L04AX05 pirfenidone, L04AX06 pomalidomide, L04AX07 dimethyl fumarate, L04AX08 darvadstrocel, L04AX09 diroximel fumarate; or any combination thereof.91.168061 / 017. The composition for use as claimed in any of claims 1 to 5, wherein the one or more immune modulating drugs are selected from mTOR inhibitors, preferably selected from A-443654, ABI009, AEZS-126, AEZS-127, AR-12, AR-42, AT13148, apitolisib, AZD6482, AZD8055, BGT226, BEZ235, bimiralisib, biolimus a9, CAL-101, CAL-120, CAL-263, CC-11, CC-223, GCS-0941, dactolisib, DB12180, everolimus, EXEL-2044, EXEL-3885, EXEL-4431, EXEL-7518, EZN-4150, farnesyl thioslicyclic acid, GDC- 0369, GDC-0941, gedatolisib, GSK60-693, GSK1059615, GSK1720070, INK128, Ku- 0063794, LY294002, LY3023414, LY317615, MHY1485, miltefosine, MK2206, MKC- 1, KRX-0401, KU-006374, myolimus, novolimus, NV- 128, NVP-BEZ235, omipalisib, OSI-027, P2281, palmoid 529, PF-04691502, PI-103, PP121, PP242, PP30, PQR309 PX- 866, QLT-0447, ridaforolimus, sapanisertib, SF-1126, SF2626, rapamycin, SR13668, TAE226, TAFA-93, temsirolimus, Torin 1, Torin 2, TOP216, vistusertib, voxtalisib, VQD-002, WAY-600, WJD008, wortmannin, WX-037, WYE-354, WYE-687, WYE- 125132, XL147, XL388, XL-765 YM-58483, zotarolimus and 3BDO or derivatives thereof, more preferably wherein the one or more immune modulating drugs are selected from rapamycin, everolimus, temsirolimus or derivatives thereof, even more preferably wherein the one or more immune modulating drugs are selected from rapamycin or derivates thereof.

8. The composition for use as claimed in any of claims 1 to 7, wherein the one or more immune modulating drugs are in the form of a cyclodextrin complex.

9. The composition for use as claimed in any of claims 1 to 8, wherein the silicon nanoparticles comprise at least 50 wt% silicon with an oxidation level of zero, even more preferably wherein said silicon nanoparticles comprise at least 80 wt% silicon with an oxidation level of zero.

10. The composition for use as claimed in any one of claims 1 to 9, wherein said use comprises administration of the composition prior to, simultaneously with or after said one or more immunogenic drugs.91.168061 / 0111. The composition for use as claimed in claim 10, wherein the composition is administered within 24 hours after the administration of said one or more immunogenic drugs12. The composition for use as claimed in any of claims 1 to 11, wherein said composition is formulated for parental administration.

13. Use of a composition as defined by any one of claims 1 to 12 for the manufacture of a medicament for the treatment or prevention of one or more unwanted immune responses in a subject, wherein the one or more unwanted immune responses are caused by the use of one or more immunogenic drugs and wherein the composition comprises negatively charged non-polymeric nanoparticles.

14. Use of a composition as defined by any one of claims 1 to 12 in the treatment or prevention of one or more unwanted immune responses in a subject, wherein the one or more unwanted immune responses are caused by the use of one or more immunogenic drugs and wherein the composition comprises negatively charged non-polymeric nanoparticles.

15. A method of treating or preventing one or more unwanted immune responses in a subject, wherein the one or more unwanted immune responses are caused by the use of one or more immunogenic drugs, wherein the method comprises the administration of a composition as defined by any one of claims 1 to 12.

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