US 20150258127A1 (19) United States (12) Patent Application Publication (10) Pub. No.: US 2015/0258127 A1 Terzi et al. (43) Pub. Date: Sep. 17, 2015 (54) METHODS FOR PREVENTING A613 L/496 (2006.01) ANTIPHOSPHOLIPID SYNDROME (APS) A613 L/4745 (2006.01) A6II 45/06 (2006.01) (71) Applicants: INSERM (INSTITUT NATIONAL DE A613 L/45 (2006.01) LA SANTE ET DE LA RECHERCHE A613 L/4439 (2006.01) MEDICALE), Paris (FR): A613 L/436 (2006.01) UNIVERSITE PARIS DESCARTES, A63/675 (2006.01) Paris (FR) A 6LX3/59 (2006.01) A 6LX3/5377 (2006.01) (72) Inventors: Fabiola Terzi, Paris (FR); Guillaume A613 L/437 (2006.01) Canaud, Paris (FR); Frank Bienaime, (52) U.S. Cl. Paris (FR) CPC ........... A6 IK3I/685 (2013.01); A61 K3I/5377 (2013.01); A61 K3I/635 (2013.01); A61 K (21) Appl. No.: 14/439,266 3 1/496 (2013.01); A61 K3I/4745 (2013.01): A6 IK3I/437 (2013.01); A61 K3I/415 (22) PCT Filed: Oct. 31, 2013 (2013.01); A61 K3I/4439 (2013.01); A61 K PCT/EP2013/072840 31/436 (2013.01); A61 K3I/675 (2013.01); (86). PCT No.: A6 IK3I/519 (2013.01); A61K 45/06 (2013.01) S371 (c)(1), (2) Date: Apr. 29, 2015 (57) ABSTRACT The present invention relates to the prevention or treatment of (30) Foreign Application Priority Data antiphospholipid syndrome (APS) in a patient in need thereof (e.g. patients affected with primary APS, a secondary APS, a Oct. 31, 2012 (EP) .................................. 12306365.3 catastrophic APS (CAPS) or a transplant recipient with antiphospholipid antibodies (APA)). The present invention Publication Classification also relates to the prevention APS-related vascular lesions in said a patient in need thereof. The present invention further (51) Int. Cl. relates to PI3K-AKT-mTOR pathway inhibitor for use in A6 IK3I/685 (2006.01) inhibiting endothelial mTORC activation triggered by APA A6 IK3I/635 (2006.01) in a patient in need thereof. Patent Application Publication Sep. 17, 2015 Sheet 1 of 4 US 2015/0258127 A1 5 5 SLE SLE Control APS APS APS 15. 10 SLE SLE Control APS- APS- APS 20 1 s 510 O SLE SLE Control APS- APS- APS Figure 1 Patent Application Publication Sep. 17, 2015 Sheet 2 of 4 US 2015/0258127 A1 P-AKT (Ser473) -> P-AKT (Thr308) -> B Actin-D 3. S. 3. S s s . e s 2 sa. C g sa R n e t op H gNa n NH IgG APA IgG Vehicle Vehicle PP242 LY294.002 Siro 1 Siro 48h P-AKT (Ser79) -> - 60kDa Total AKT - 60kDa P-S6RP - 32kDa S6RP - 32kDa Figure 2 Patent Application Publication Sep. 17, 2015 Sheet 3 of 4 US 2015/0258127 A1 Go S 9S. E. Ais 9 is to S TXAPA- Siro- Siro + TXAPA TXAPA- Siro- Siro- TXAPA- Siro- Siro + TXAPA TxAPA fiti TxAPA- Siro- Siro- TXAPA- Siro- Siro + TxAPA- TxAPA C D 5 n S. E E S 5 ? L s c E O a. TXAPA- Siro- Siro + o Months TXAPA Figure 3 Patent Application Publication Sep. 17, 2015 Sheet 4 of 4 US 2015/0258127 A1 A C) SE &N 25 iii. X 35 20 g 15 8 SD8 cu O 5 2 : ck ke SS o TXAPA B ifiti a Ša. 5 CS is A 9 R ck kick D 8x8 3.x: TXAPA- Siro- Siro + TXAPA C es SQ 9.se E 9. 3.O 5 St. ck kick us e TXAPA- Siro- Siro TAPA Figure 4 US 2015/0258127 A1 Sep. 17, 2015 METHODS FOR PREVENTING vascular alterations in APS. The pathophysiology of these ANTIPHOSPHOLIPID SYNDROME (APS) lesions is unknown and efficient therapeutic strategy are lack 1ng. FIELD OF THE INVENTION 0006 mTORC kinase is a central node signalling path ways that regulate cellular growth, proliferation and Survival. 0001. The present invention relates to the prevention or mTOR is a component of two functionally distinct com treatment of antiphospholipid syndrome (APS). The present plexes. mTOR complex 1 (mTORC1) stimulates ribosome invention also relates to the prevention APS-related vascular biogenesis and protein translation by phosphorylating S6 lesions in a patient in need thereof (e.g. patients affected with kinase while in turn activates S6 ribosomal protein (S6RP), primary APS, a secondary APS, a catastrophic APS (CAPS) and 4E-BP1 protein (4EBP1). mTOR complex 2 (mTORC2) or a transplant recipient with antiphospholipid antibodies promotes Survival, proliferation or migration depending on (APA)). The present invention further relates to the inhibition the cellular context, through AKT phosphorylation on Ser'. of endothelial mTORC activation triggered by APA in a An important and complex cross-regulation exists between patient in need thereof. mTORC1 and mTORC2. Indeed, the activation of AKT by mTORC2 stimulates mTORC1, whereas mTORC1 reduces BACKGROUND OF THE INVENTION mTORC2 activation'. mTORC has been shown to play an important role in the vascular narrowing secondary to 0002 Antiphospholipid syndrome (APS) is an autoim mechanical endothelial injury in both experimental models mune disease characterized by the presence of circulating and patients undergoing arterial angioplasty notably by pro antiphospholipid antibodies (APA also referred as aPL) that moting vascular smooth muscle cells (VSMC) proliferation cause arterial, venous and Small vessels thrombosis and/or in the media'. Indeed the mTORC inhibitor sirolimus is obstetrical complications consisting in pregnancy loss or pre now currently used to prevent reactive arterial stenosis after term birth due to pre-eclampsia or placental insufficiency1. coronary artery stenting. APA are a family of autoantibodies that recognize various 0007. However, the activation of the mTOR pathway in phospholipids and plasma proteins with affinity for anionic endothelial cells by APA leading to the vascular lesions of cell surface phospholipids. There are three main types of APA: lupus anticoagulant (LA), anti-cardiolipin (aCL) and APSN has never been studied nor even suggested until now. anti-f2 glycoprotein I antibodies (anti-B2GPI)". APS is SUMMARY OF THE INVENTION observed either isolated or in association with in a number of autoimmune disorders, i.e. systemic lupus erythematosus 0008. In a first aspect, the present invention also relates to (SLE). a PI3K-AKT-mTOR pathway inhibitor for use in the preven 0003 APS is considered as the most frequent cause for tion of APS-related vascular lesions in a patient in need acquired thrombophilia and is associated with high morbidity thereof. and mortality'. APS account for 20% of the stroke in young 0009. In a second aspect, the present invention also relates patients. In addition, APS represents a major adverse prog to a PI3K-AKT-mTOR pathway inhibitor for use in inhibiting nostic factor inpatients with SLE. The main consequence of endothelial mTORC activation triggered antiphospholipid the APS is thrombotic complications, and so far, the only antibodies (APA) in a patient in need thereof. treatment, which has been shown to reduce the vascular com 0010. In a third aspect, the present invention further relates plications in APS patients, is permanent anticoagulation. to a pharmaceutical composition for use in the prevention of However, this regimen does not completely prevent the recur APS-related vascular lesions comprising a PI3K-AKT rence of thrombosis in high risks patients and is associated mTOR pathway inhibitor and a pharmaceutically acceptable with an increase incidence of bleeding. carrier. 0004 Although thrombosis is considered as the key fea 0011. In still another aspect, the present invention relates ture of the vascular disease in APS, chronic arterial and arte to a kit comprising at least two PI3K-AKT-mTOR pathway riolar lesions have been frequently associated. These lesions inhibitors, as a combined preparation for simultaneous, sepa consist mainly in thickening of the intima and the media and rate or sequential use in the prevention of APS-related vascu are often associated with increased cellularity of the two lar lesions. layers'''. These lesions have been particularly well charac terized in the kidney and called APS-nephropathy (APSN). DETAILED DESCRIPTION OF THE INVENTION These vascular changes lead to progressive fibrosis that ulti 0012. The present invention is based on the vascular acti mately results in end-stage renal failure (ESRF)'''. More vation of both mTORC1 and 2 pathways in APSN as well as over, it has been reported that kidney transplant recipients in others critical arterial beds in patients with severe APS. with APA are at greater risk to develop thrombotic complica Remarkably, this activation concerned selectively the endot tion'''. In addition to thrombotic complication, it has been helial cells but correlated with proliferation of both endothe observed that these patients developed typical features of lial and Smooth muscle cells and, more importantly, with APSN recurrence on the allograft'. These lesions led to a fast vascular lesions. Thus, the inventors demonstrated for the first decline of the measured glomerular filtration rate (mGFR). time the crucial role played by endothelial mTORC pathway 0005 To date the effort made to elucidate the pathogenesis activation in the development of the fibrous intimal hyperpla of APS have focused on the mechanisms of thrombosis for sia in APS patients. Mechanistically purified IgG from mation whereas the pathophysiological processes respon patients with APS activate both mTORC1 and mTORC2 in sible for the chronic vascular changes associated with APS cultured endothelial cells in a complement independent man have not been investigated. In that regard, a better understand ner. Briefly, these antiphospholipid IgG were collected from ing of APSN pathogeny could represent an important mile 12 different patients. Among these patients, 7 underwent a stone to elaborate therapeutic strategies limiting the chronic kidney biopsy that revealed the presence of APS nephropathy US 2015/0258127 A1 Sep. 17, 2015 with the characteristic vascular lesions. All the tested 0019. As used herein, the term “patient” refers to an ani antiphospholipid IgG were able to activate the mTORC path mal, preferably to a mammal, even more preferably to a way in vitro and in Vivo and the intensity of activation corre human, including adult and child.